KR20240032915A - Tricyclic compounds as inhibitors of KRAS - Google Patents

Abstract

Disclosed are compounds of Formula I, methods of using the compounds to inhibit KRAS activity, and pharmaceutical compositions comprising such compounds. The compounds are useful for treating, preventing or ameliorating diseases or disorders associated with KRAS activity, such as cancer.

Description

Tricyclic compounds as inhibitors of KRAS

Related applications

This application relates to U.S. Provisional Patent Application No. 63/219,274, filed on July 7, 2021; U.S. Provisional Application No. 63/292,774, filed December 22, 2021; and U.S. Provisional Application No. 63/310,811 filed on February 16, 2022, the contents of which are incorporated in their entirety.

field of invention

The present disclosure provides compounds as well as compositions and methods of use thereof. This compound modulates KRAS activity and is useful in the treatment of various diseases, including cancer.

Ras proteins are part of a family of small GTPases that are activated by growth factors and various extracellular stimuli. The Ras family regulates intracellular signaling pathways responsible for cell growth, migration, survival, and differentiation. Activation of RAS proteins at the cell membrane results in the binding of key effectors and initiation of a cascade of intracellular signaling pathways within the cell, including the RAF and PI3K kinase pathways. Somatic mutations in RAS can lead to uncontrolled cell growth and malignant transformation, while activation of RAS proteins is tightly regulated in normal cells (Simanshu, D. et al. Cell 170.1 (2017):17-33).

The Ras family includes three members: KRAS, NRAS, and HRAS. RAS mutant cancers account for approximately 25% of human cancers. KRAS is the most frequently mutated isoform, accounting for 85% of all RAS mutants, while NRAS and HRAS are found to be mutated in 12% and 3% of all Ras mutant cancers, respectively (Simanshu, D. et al. Cell 170.1 (2017):17-33). KRAS mutations are prevalent among the top three most lethal cancer types: pancreatic cancer (97%), colorectal cancer (44%), and lung cancer (30%) (Cox, A.D. et al. Nat Rev Drug Discov (2014) 13: 828-51). Most RAS mutations occur at amino acid residues 12, 13, and 61. The frequency of specific mutations varies between RAS gene isoforms, but G12 and Q61 mutations are predominant in KRAS and NRAS, respectively, and G12, G13, and Q61 mutations are most frequent in HRAS. Moreover, the spectrum of mutations within RAS isoforms differs between cancer types. For example, KRAS G12D mutations predominate in pancreatic cancer (51%) followed by colorectal adenocarcinoma (45%) and lung cancer (17%), whereas KRAS G12 V mutations predominate in pancreatic cancer (30%) followed by colorectal adenocarcinoma (27 %) and lung adenocarcinoma (23%) (Cox, A.D. et al. Nat Rev Drug Discov (2014) 13:828-51). In contrast, KRAS G12C mutations are prevalent in non-small cell lung cancer (NSCLC), including 11 to 16% of lung adenocarcinomas and 2 to 5% of pancreatic and colorectal adenocarcinomas (Cox, A.D. et al. Nat. Rev. Drug Discov. (2014) 13:828-51). Genomic studies across hundreds of cancer cell lines have demonstrated that cancer cells with KRAS mutations are highly dependent on KRAS function for cell growth and survival (McDonald, R. et al. Cell 170 (2017): 577-592). The role of mutant KRAS as an oncogenic driver has been demonstrated by intensive in vivo experimental evidence showing that mutant KRAS is required for early tumor onset and maintenance in animal models (Cox , A.D. et al. Nat Rev Drug Discov (2014) 13:828-51).

Taken together, these findings suggest that KRAS mutations play an important role in human cancer; Therefore, the development of inhibitors targeting mutant KRAS may be useful in the clinical treatment of diseases characterized by KRAS mutations.

The present disclosure particularly relates to compounds of formula (I):

or a pharmaceutically acceptable salt thereof, wherein the constituent variables are defined herein.

The disclosure further provides pharmaceutical compositions comprising a compound of the disclosure, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient.

The present disclosure further includes a method of inhibiting KRAS activity, the method comprising administering to the subject a compound of the disclosure or a pharmaceutically acceptable salt thereof. The disclosure also provides for the use of the compounds described herein in the manufacture of a medicament for use in therapy. The disclosure also provides compounds described herein for use in therapy.

The disclosure further provides a method of treating a disease or disorder in a patient comprising administering a therapeutically effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof.

compound

In one aspect, provided herein is a compound having the formula (I):

During the ceremony:

Y is N or CH;

R ¹ is selected from Cl, CH ₃ , CH ₂ F, CHF ₂ and CF ₃ ;

Cy ¹ is selected from:

R ² is selected from F and Cl;

R ³ is selected from:

and

Cy ² is selected from:

However, the compounds of formula I are other than:

In one embodiment of Formula I, or a pharmaceutically acceptable salt thereof,

Y is N or CH;

R ¹ is selected from Cl, CH ₃ , CH ₂ F, CHF ₂ and CF ₃ ;

Cy ¹ is selected from:

R ² is selected from F and Cl;

R ³ is selected from:

and

Cy ² is selected from:

However, the compounds of formula I are other than:

In another embodiment, the compound of Formula I is a compound of Formula II:

or a pharmaceutically acceptable salt thereof, in the formula:

R ¹ is selected from Cl and CH ₃ ;

Cy ¹ is selected from:

R ³ is selected from:

and

Cy ² is selected from:

In one embodiment of Formula I, or a pharmaceutically acceptable salt thereof,

Y is N or CH;

R ¹ is selected from Cl, CH ₃ , CH ₂ F, CHF ₂ and CF ₃ ;

Cy ¹ is selected from:

R ² is selected from F and Cl;

R ³ is selected from:

and

Cy ² is selected from:

In one embodiment, Y is CH. In one embodiment Y is N.

In one embodiment, Cy ¹ is Cy ¹ -c, Cy ¹ -l, Cy ¹ -m, Cy ¹ -n, Cy ¹ -o, Cy ¹ -p, Cy ¹ -q, Cy ¹ -r, Cy ¹ -s and Cy ¹ -t. In one embodiment, Cy ¹ is Cy ¹ -l, Cy ¹ -m, Cy ¹ -n, Cy ¹ -o, Cy ¹ -p, Cy ¹ -q, Cy ¹ -r, Cy ¹ -s and Cy ¹ Selected from -t. In one embodiment, Cy ¹ is Cy ¹ -c, Cy ¹ -m, Cy ¹ -n, Cy ¹ -o, Cy ¹ -p, Cy ¹ -q, Cy ¹ -r, Cy ¹ -s and Cy ¹ Selected from -t. In one embodiment, Cy ¹ is selected from Cy ¹ -f, Cy ¹ -g, Cy ¹ -h, Cy ¹ -i, Cy ¹ -j, Cy ¹ -k and Cy ¹ -l. In one embodiment, Cy ¹ is selected from Cy ¹ -a, Cy ¹ -m, Cy ¹ -n, Cy ¹ -o, Cy ¹ -p and Cy ¹ -q. In one embodiment, Cy ¹ is selected from Cy ¹ -c, Cy ¹ -d, Cy ¹ -e, Cy ¹ -r, Cy ¹ -s and Cy ¹ -t.

In one embodiment, Cy ¹ is selected from Cy ¹ -a, Cy ¹ -c and Cy ¹ -r. In one embodiment, Cy ¹ is selected from Cy ¹ -c and Cy ¹ -r. In one embodiment, Cy ¹ is selected from Cy ¹ -s and Cy ¹ -t.

In one embodiment, Cy ¹ is Cy ¹ -l. In one embodiment, Cy ¹ is Cy ¹ -m. In one embodiment, Cy ¹ is Cy ¹ -n. In one embodiment, Cy ¹ is Cy ¹ -o. In one embodiment, Cy ¹ is Cy ¹ -p. In one embodiment, Cy ¹ is Cy ¹ -q. In one embodiment, Cy ¹ is Cy ¹ -r. In one embodiment, Cy ¹ is Cy ¹ -s. In one embodiment, Cy ¹ is Cy ¹ -t.

In one embodiment, Cy ¹ is Cy ¹ -a, Cy ¹ -b, Cy ¹ -c, Cy ¹ -d, Cy ¹ -e, Cy ¹ -f, Cy ¹ -g, Cy ¹ -i and Cy ¹ Selected from -j. In one embodiment, Cy ¹ is selected from Cy ¹ -a, Cy ¹ -b, Cy ¹ -c, Cy ¹ -d and Cy ¹ -e. In one embodiment, Cy ¹ is selected from Cy ¹ -f, Cy ¹ -g, Cy ¹ -h, Cy ¹ -i, Cy ¹ -j and Cy ¹ -k. In one embodiment, Cy ¹ is selected from Cy ¹ -c, Cy ¹ -d and Cy ¹ -e. In one embodiment, Cy ¹ is selected from Cy ¹ -a, Cy ¹ -b and Cy ¹ -k. In one embodiment, Cy ¹ is selected from Cy ¹ -a and Cy ¹ -b. In one embodiment, Cy ¹ is selected from Cy ¹ -h and Cy ¹ -k.

In one embodiment, Cy ¹ is Cy ¹ -a. In one embodiment, Cy ¹ is Cy ¹ -b. In one embodiment, Cy ¹ is Cy ¹ -c. In one embodiment, Cy ¹ is Cy ¹ -d. In one embodiment, Cy ¹ is Cy ¹ -e. In one embodiment, Cy ¹ is Cy ¹ -f. In one embodiment, Cy ¹ is Cy ¹ -g. In one embodiment, Cy ¹ is Cy ¹ -h. In one embodiment, Cy ¹ is Cy ¹ -i. In one embodiment, Cy ¹ is Cy ¹ -j. In one embodiment, Cy ¹ is Cy ¹ -k.

In one embodiment, R ¹ is selected from CH ₃ , CH ₂ F, CHF ₂ and CF ₃ . In one embodiment, R ¹ is selected from Cl, CH ₃ , CHF ₂ and CF ₃ . In one embodiment, R ¹ is selected from CH ₂ F, CHF ₂ and CF ₃ . In one embodiment, R ¹ is selected from Cl, CH ₂ F, CHF ₂ and CF ₃ . In one embodiment, R ¹ is selected from Cl and CH ₃ . In one embodiment, R ¹ is selected from Cl and CF ₃ . In one embodiment, R ¹ is selected from CH ₃ and CF ₃ . In one embodiment, R ¹ is Cl. In one embodiment, R ¹ is CH ₂ F. In one embodiment, R ¹ is CHF ₂ . In one embodiment, R ¹ is CH ₃ . In one embodiment, R ¹ is CF ₃ .

In one embodiment, R ² is F. In one embodiment R ² is Cl.

In one embodiment, R ³ is selected from R ³ -a and R ³ -b. In one embodiment, R ³ is selected from R ³ -b and R ³ -c. In one embodiment, R ³ is selected from R ³ -a and R ³ -c. In one embodiment, R ³ is R ³ -a. In one embodiment, R ³ is R ³ -b. In one embodiment, R ³ is R ³ -c.

In one embodiment, R ³ is selected from R ³ -b, R ³ -c and R ³ -d. In one embodiment, R ³ is selected from R ³ -b and R ³ -d. In one embodiment, R ³ is selected from R ³ -c and R ³ -d.

In one embodiment, R ³ is selected from R ³ -a and R ³ -d. In one embodiment, R ³ is R ³ -d.

In one embodiment, Cy ² is selected from Cy ² -b, Cy ² -d, Cy ² -e and Cy ² -f. In one embodiment, Cy ² is selected from Cy ² -b and Cy ² -d. In one embodiment, Cy ² is selected from Cy ² -c and Cy ² -d.

In one embodiment, Cy ² is selected from Cy ² -a, Cy ² -c and Cy ² -d. In one embodiment, Cy ² is selected from Cy ² -a, Cy ² -b and Cy ² -d. In one embodiment, Cy ² is selected from Cy ² -a, Cy ² -b and Cy ² -c. In one embodiment, Cy ² is selected from Cy ² -d, Cy ² -e and Cy ² -f. In one embodiment, Cy ² is selected from Cy ² -a and Cy ² -b. In one embodiment, Cy ² is selected from Cy ² -c and Cy ² -f. In one embodiment, Cy ² is selected from Cy ² -e and Cy ² -f.

In one embodiment, Cy ² is Cy ² -a. In one embodiment, Cy ² is Cy ² -b. In one embodiment, Cy ² is Cy ² -c. In one embodiment, Cy ² is Cy ² -d. In one embodiment, Cy ² is Cy ² -e. In one embodiment, Cy ² is Cy ² -f.

In another embodiment, the compound of Formula I is other than:

In one embodiment, the compound of Formula I is other than:

In another embodiment, a compound of formula herein is a compound of formula or a pharmaceutically acceptable salt thereof.

In one embodiment, the compound of Formula I is selected from:

2-((2 S ,4 S )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinoline- 8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4- fluorobut-2-enoyl)piperidin-2-yl)acetonitrile;

2-((2 S ,4 S )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(2-methoxy-3 -methylphenyl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4-fluo Robut-2-enoyl)piperidin-2-yl)acetonitrile;

2-((2 S ,4 S )-4-(7-(3-chloro-2-methoxyphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)- 6-fluoro-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4- fluorobut-2-enoyl)piperidin-2-yl)acetonitrile;

2-(( 2S , 4S )-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-(( S )-1-(( S )-1-methyl Pyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1- (( E )-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile;

1-(4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-(( S )-1-(( S )-1-methylpyrrolidin-2-yl) Ethoxy)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidin-1-yl)prop- 2-en-1-one;

2-((2 S ,4 S )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluo Ro-8-methyl- 1H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4-methoxybut-2-enoyl )piperidin-2-yl)acetonitrile;

2-(( 2S , 4S )-4-(6-fluoro-8-methyl-7-(1-methyl- 1H -indazol-6-yl)-4-(( S )-1- (( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-( ( E )-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile;

2-(( 2S , 4S )-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-(( S )-1-(( S )- 1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4 -fluorobut-2-enoyl)piperidin-2-yl)acetonitrile;

2-(( 2S , 4S )-4-(6-fluoro-8-methyl-7-(1-methyl- 1H -indazol-3-yl)-4-(( S )-1- (( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-( ( E )-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile;

2-(( 2S , 4S )-4-(6-fluoro-7-(4-fluorophenyl)-8-methyl-4-(( S )-1-(( S )-1-methyl Pyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4-methoxy but-2-enoyl)piperidin-2-yl)acetonitrile;

8-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-(( S )-1-(( S )-1-methylpyrrolidin-2 -yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)-1-naphtonitrile;

2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro -8-methyl-1H-imidazo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)aceto nitrile;

2-((2 S ,4 S )-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-(( S )-1-(( S )-1 -methylpyrrolidin-2-yl)ethoxy)-1 H -imidazo[4,5- c ]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidine-2- 1) Acetonitrile;

8-(1-((2 R ,4 S )-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-(( S )-1- (( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)-1-naph Tonitrile;

2-((2 S ,4 S )-4-(7-(5,6-dimethyl-1 H -indazol-4-yl)-4-(3-(ethyl(methyl)amino)azetidine- 1-yl)-6-fluoro-8-methyl-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(2-fluoroacrylo 1) piperidin-2-yl) acetonitrile;

8-(6-fluoro-1-(1-(( E )-4-fluorobut-2-enoyl)piperidin-4-yl)-8-methyl-4-(( S )-1 -(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)-1- naphtonitrile;

8-(1-((2 S ,4 S )-2-(cyanomethyl)-1-(2-fluoroacryloyl)piperidin-4-yl)-4-(3-(dimethyl amino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)- 1-naphtonitrile;

2-((2 S ,4 S )-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoro Quinolin-8-yl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4-methoxybut-2-ene oil) piperidin-2-yl) acetonitrile;

2-((2 S ,4 S )-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoro Quinolin-8-yl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4-fluorobut-2-ene oil) piperidin-2-yl) acetonitrile; and

2-((2 S ,4 S )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinoline- 8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4- methoxybut-2-enoyl)piperidin-2-yl)acetonitrile;

Or a pharmaceutically acceptable salt thereof.

In another embodiment, the compound of Formula I is selected from:

2-((2 S ,4 S )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinoline- 8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4- fluorobut-2-enoyl)piperidin-2-yl)acetonitrile;

2-((2 S ,4 S )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(2-methoxy-3 -methylphenyl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4-fluo Robut-2-enoyl)piperidin-2-yl)acetonitrile;

2-((2 S ,4 S )-4-(7-(3-chloro-2-methoxyphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)- 6-fluoro-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4- fluorobut-2-enoyl)piperidin-2-yl)acetonitrile;

2-(( 2S , 4S )-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-(( S )-1-(( S )-1-methyl Pyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1- (( E )-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile;

1-(4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-(( S )-1-(( S )-1-methylpyrrolidin-2-yl) Ethoxy)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidin-1-yl)prop- 2-en-1-one;

2-(( 2S , 4S )-4-(6-fluoro-8-methyl-7-(1-methyl- 1H -indazol-6-yl)-4-(( S )-1- (( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-( ( E )-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile;

2-(( 2S , 4S )-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-(( S )-1-(( S )- 1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4 -fluorobut-2-enoyl)piperidin-2-yl)acetonitrile;

2-(( 2S , 4S )-4-(6-fluoro-8-methyl-7-(1-methyl- 1H -indazol-3-yl)-4-(( S )-1- (( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-( ( E )-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile;

2-(( 2S , 4S )-4-(6-fluoro-7-(4-fluorophenyl)-8-methyl-4-(( S )-1-(( S )-1-methyl Pyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4-methoxy but-2-enoyl)piperidin-2-yl)acetonitrile;

8-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-(( S )-1-(( S )-1-methylpyrrolidin-2 -yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)-1-naphtonitrile;

8-(1-((2 R ,4 S )-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-(( S )-1- (( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)-1-naph tonitrile;

2-((2 S ,4 S )-4-(7-(5,6-dimethyl-1 H -indazol-4-yl)-4-(3-(ethyl(methyl)amino)azetidine- 1-yl)-6-fluoro-8-methyl-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(2-fluoroacrylo 1) piperidin-2-yl) acetonitrile;

8-(6-fluoro-1-(1-(( E )-4-fluorobut-2-enoyl)piperidin-4-yl)-8-methyl-4-(( S )-1 -(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)-1- naphtonitrile;

8-(1-((2 S ,4 S )-2-(cyanomethyl)-1-(2-fluoroacryloyl)piperidin-4-yl)-4-(3-(dimethyl amino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)- 1-naphtonitrile;

2-((2 S ,4 S )-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoro Quinolin-8-yl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4-methoxybut-2-ene oil) piperidin-2-yl) acetonitrile;

2-((2 S ,4 S )-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoro Quinolin-8-yl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4-fluorobut-2-ene oil) piperidin-2-yl) acetonitrile; and

2-((2 S ,4 S )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinoline- 8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4- methoxybut-2-enoyl)piperidin-2-yl)acetonitrile;

Or a pharmaceutically acceptable salt thereof.

In another embodiment, the compound of Formula I is selected from:

2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-((S)- 1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4- methoxybut-2-enoyl)piperidin-2-yl)acetonitrile;

2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-((S)- 1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacrylo 1) piperidin-2-yl) acetonitrile;

2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1 -methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl )piperidin-2-yl)acetonitrile;

2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1 -methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-meth Toxybut-2-enoyl)piperidin-2-yl)acetonitrile;

2-((2S,4S)-4-(6-fluoro-7-(7-fluoro-2-methylquinolin-8-yl)-8-methyl-4-((S)-1-(( S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluo Roacryloyl)piperidin-2-yl)acetonitrile;

2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methylisoquinolin-4-yl)-4-((S)-1-((S)-1 -methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-meth Toxybut-2-enoyl)piperidin-2-yl)acetonitrile;

2-((2S,4S)-4-(6-fluoro-7-(7-fluoroquinolin-8-yl)-8-methyl-4-((S)-1-((S)-1 -methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl )piperidin-2-yl)acetonitrile;

2-(4-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1 -((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl -1H-indazol-3-yl)acetonitrile;

2-((2S,4S)-4-(4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5 -methyl-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoro Acryloyl)piperidin-2-yl)acetonitrile;

2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidine -1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacrylo 1) piperidin-2-yl) acetonitrile;

2-(4-(1-((2 R ,4 S )-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-(( S ) -1-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)- 1-methyl-1 H -indol-3-yl)acetonitrile;

2-(4-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-(( S )-1-(( S )-1-methylpyrroli din-2-yl) ethoxy) -1 H -[1,2,3] triazolo [4,5- c ] quinolin-7-yl) -1-methyl-1 H -indol-3-yl) acetonitrile;

2-((2S,4S)-4-(8-chloro-7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3 -methylazetidin-1-yl)-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacrylo 1) piperidin-2-yl) acetonitrile;

2-((2S,4S)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-8-methyl-4-((S)-1-((S)- 1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacrylo 1) piperidin-2-yl) acetonitrile; and

2-((2S,4S)-4-(8-chloro-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6- Fluoro-5-methyl-1H-indazol-4-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoro Acryloyl)piperidin-2-yl)acetonitrile;

Or a pharmaceutically acceptable salt thereof.

In one embodiment, the compound of Formula I is selected from:

2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro -8-methyl-1H-imidazo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)aceto nitrile;

2-((2 S ,4 S )-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-(( S )-1-(( S )-1 -methylpyrrolidin-2-yl)ethoxy)-1 H -imidazo[4,5- c ]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidine-2- 1) Acetonitrile;

Or a pharmaceutically acceptable salt thereof.

For the sake of clarity, it is further recognized that certain features of the invention that are described in the context of separate embodiments may also be provided in combination in a single embodiment (although embodiments may also be provided in multiple dependent forms). are intended to be combined together). Conversely, for the sake of brevity, various features of the invention that are described in the context of a single embodiment may also be provided separately or in any suitable subcombination. Accordingly, it is contemplated that the features described as embodiments of compounds of formula (I) may be combined in any suitable combination.

Compounds described herein may be asymmetric (eg, having more than one stereogenic center). All stereoisomers, including enantiomers and diastereomers, are intended unless otherwise indicated. Compounds of the invention containing asymmetrically substituted carbon atoms may be isolated in optically active or random forms. Methods for preparing optically active forms from optically inactive starting materials, such as by resolution of racemic mixtures or by stereoselective synthesis, are known in the art. Many geometric isomers of olefins, such as C=N double bonds, etc., may exist in the compounds described herein, and all such stable isomers are contemplated in the present invention. Cis and trans geometric isomers of the compounds of the invention are described and can be isolated as mixtures of isomers or as separate isomeric forms.

Resolution of racemic mixtures of compounds can be accomplished by any of many methods known in the art. One method involves fractional recrystallization using chiral splitting acids, which are optically active salt-forming organic acids. Suitable resolving agents for the fractional recrystallization process include, for example, optically active acids such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, the D and L forms of lactic acid, and various optically active camphorsulfonic acids, For example, β-camphorsulfonic acid. Other resolving agents suitable for fractional crystallization methods include α-methylbenzylamine (e.g. in S and R forms or in diastereomerically pure form), 2-phenylglycinol, norephedrine, ephedrine, N -methylephedrine, cyclohexyl Includes stereomerically pure forms of ethylamine, 1,2-diaminocyclohexane, etc.

Resolution of racemic mixtures can also be performed by elution on a column filled with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). Suitable elution solvent compositions can be determined by one skilled in the art.

In some embodiments, the compounds of the invention have the ( R )-configuration. In another embodiment, the compound has the ( S )-configuration. In compounds having more than one chiral center, each of the chiral centers in the compound can independently be ( R ) or ( S ), unless otherwise indicated.

The compounds of the present invention include tautomeric forms. The tautomeric forms result from the swapping of a single bond with an adjacent double bond with simultaneous movement of a proton. Tautomeric forms include protic tautomers, which are isomeric proton addition states with the same empirical formula and total charge. Exemplary protic tautomers include ketone-enol pairs, amide-imide acid pairs, lactam-lactam pairs, enamine-imine pairs, and cyclic forms in which the proton can occupy two or more positions in the heterocyclic system, such as , 1H- and 3H-imidazole, 1H-, 2H- and 4H- 1,2,4-triazole, 1H- and 2H- isoindole and 1H- and 2H-pyrazole. Tautomeric forms can be sterically locked or in equilibrium in one form by appropriate substitution.

The compounds of the present invention may contain all isotopes of atoms present in the intermediate or final compound. Isotopes include atoms with the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. One or more constituent atoms of the compounds of the present invention may be replaced or substituted with isotopes of the atom in natural or non-natural abundance. In some embodiments, the compound includes at least one deuterium atom. For example, one or more hydrogen atoms in a compound of the present disclosure may be replaced or substituted by deuterium. In some embodiments, the compound contains two or more deuterium atoms. In some embodiments, the compound contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 deuterium atoms. Synthetic methods for incorporating isotopes into organic compounds are known in the art (Deuterium Labeling in Organic Chemistry by Alan F. Thomas (New York, N.Y., Appleton-Century-Crofts, 1971; The Renaissance of H/D Exchange by Jens Atzrodt, Volker Derdau, Thorsten Fey and Jochen Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765; The Organic Chemistry of Isotopic Labeling by James R. Hanson, Royal Society of Chemistry, 2011). The compounds can be used in a variety of studies such as NMR spectroscopy, metabolic experiments and/or assays.

Substitution with heavy isotopes, such as deuterium, may provide certain therapeutic advantages due to greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and are therefore desirable in some situations. can do. (A. Kerekes et.al. J. Med. Chem. 2011 , 54 , 201-210; R. Xu et.al. J. Label Compd. Radiopharm. 2015 , 58 , 308-312).

The term “compound” as used herein is meant to include all stereoisomers, geometric isomers, tautomers and isotopes of the depicted structure. The term is also meant to refer to compounds of the invention, regardless of how they are made, e.g. synthetically, through biological processes (e.g. metabolism or enzymatic transformation) or combinations thereof. .

All compounds, and pharmaceutically acceptable salts thereof, may be isolated or may be found with other substances, such as water or solvents (e.g., hydrates and solvates). When in the solid state, the compounds described herein or salts thereof may exist in various forms, such as solvates, including hydrates. Since the compound may be in any solid state form, such as a polymorph or solvate, unless clearly indicated otherwise, references herein to the compound and its salts are intended to encompass any solid state form of the compound. It must be understood.

In some embodiments, the compounds of the invention, or salts thereof, are substantially isolated. “Substantially isolated” means that a compound or salt is at least partially or substantially separated from the environment in which it was formed or detected. Partial isolation may include, for example, compositions enriched in compounds of the invention. Substantial separation is at least about 50% by weight, at least about 60% by weight, at least about 70% by weight, at least about 80% by weight, at least about 90% by weight, at least about 95% by weight, at least about 97% by weight, or at least about 99% by weight. % of a compound of the present invention or a salt thereof.

The phrase "pharmaceutically acceptable" herein means that, within the scope of sound medical judgment, contact with human and animal tissue commensurate with a reasonable benefit/risk ratio, without undue toxicity, irritation, allergic reaction or other problems or complications. It is used to refer to a compound, material, composition and/or dosage form suitable for use.

As used herein, the expressions “ambient temperature” and “room temperature” are understood in the art and generally refer to a temperature, e.g., the approximate temperature of the room in which the reaction is conducted, e.g., a temperature of 20° C. to about 30° C. Phosphorus, refers to the reaction temperature.

The invention also includes pharmaceutically acceptable salts of the compounds described herein. As used herein, “pharmaceutically acceptable salt” refers to a derivative of a disclosed compound in which the parent compound is modified by converting an existing acid or base moiety to its salt form. Examples of pharmaceutically acceptable salts include mineral or organic acid salts of basic residues such as amines; Alkaline or organic salts of acidic moieties such as carboxylic acids; It includes, but is not limited to, etc. Pharmaceutically acceptable salts of the invention include, for example, non-toxic salts of the parent compounds formed from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or salt form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of both; Generally, non-aqueous media such as ether, ethyl acetate, alcohols (e.g. methanol, ethanol, iso-propanol or butanol) or acetonitrile (MeCN) are preferred. A list of suitable salts is given in Remington's Pharmaceutical Sciences , ^17th Ed., (Mack Publishing Company, Easton, 1985), p. 1418, Berge et al ., J. Pharm. Sci. , 1977 , 66 (1), 1-19 and Stahl et al ., Handbook of Pharmaceutical Salts: Properties, Selection, and Use , (Wiley, 2002). In some embodiments, the compounds described herein include N-oxide forms.

synthesis

The compounds of the present invention (including their salts) can be prepared using known organic synthesis techniques and can be synthesized according to any of a number of possible synthetic routes, such as those in the schemes below.

The reaction for preparing the compounds of the present invention can be carried out in a suitable solvent that can be easily selected by a person skilled in the field of organic synthesis. Suitable solvents may be substantially non-reactive with the starting materials (reactants), intermediates or products at the temperature at which the reaction is carried out, e.g., a temperature that may range from the solvent's freezing temperature to the solvent's boiling temperature. A given reaction may be carried out in one solvent or a mixture of more than one solvent. Depending on the particular reaction step, a solvent suitable for the particular reaction step can be selected by the skilled person.

Preparation of the compounds provided in the present invention may involve protection and deprotection of various chemical groups. The need for protection and deprotection, and selection of appropriate protecting groups, can be readily determined by those skilled in the art. The chemistry of protecting groups is described, for example, in Kocienski, Protecting Groups , (Thieme, 2007); Robertson, Protective Group Chemistry , (Oxford University Press, 2000); Smith et al ., March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure , ^6th Ed. (Wiley, 2007); Peturssion et al ., “Protecting Groups in Carbohydrate Chemistry,” J. Chem. Educ ., 1997, 74 (11), 1297; and Wuts et al ., Protective Groups in Organic Synthesis , 4th Ed., (Wiley, 2006).

The reaction can be monitored according to any suitable method known in the art. For example, product formation can be determined by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g. ¹ H or ¹³ C), infrared spectroscopy, spectrophotometry (e.g. UV-vis), mass spectrometry or by chromatographic methods, e.g. For example, it can be monitored by high performance liquid chromatography (HPLC) or thin layer chromatography (TLC).

The following reaction schemes provide general guidance regarding the preparation of the compounds of the present invention. Those skilled in the art will appreciate that the preparations shown in the schemes can be modified or optimized using general knowledge of organic chemistry to prepare the various compounds provided herein.

Scheme 1

Compounds of formula 1-18 can be prepared via the synthetic route outlined in Scheme 1. Starting material 1-1 is halogenated with a suitable reagent such as N -chloro-succinimide (NCS) to provide intermediate 1-2 (Hal is a halide such as F, Cl, Br or I). Next, compound 1-3 can be prepared by treating 1-2 with triphosgene. Intermediate 1-3 is then reacted with ester 1-4 to give nitro compound 1-5 , which is reacted with an appropriate reagent (e.g. Treatment with POCl ₃ ) can provide compounds 1-6 . Compound 1-8 can be produced by performing the S _N Ar reaction of intermediate 1-6 with amine 1-7 (PG is an appropriate protecting group such as Boc). The nitro group in 1-8 can be reduced to NH ₂ in the presence of a reducing agent (eg, acetic acid or Fe in sodium dithionite). Intermediate 1-9 is then subjected to a cyclization reaction (e.g., using triethyl orthoformate) to give intermediate 1-10 , which is then subjected to a SnAr reaction with sodium thiomethoxide to give 1-11 can be provided. 1-12 (where M is a boronic acid, boronic acid ester, or appropriately substituted metal or metalloid [e.g., M is B(OR) ₂ , Sn(Alkyl) ₃ , Zn-Hal, or CF ₃ TMS ]) and standard Suzuki cross-coupling conditions (e.g., in the presence of a palladium catalyst and an appropriate base), or standard Stille cross-coupling conditions (e.g., in the presence of a palladium catalyst), or standard Negishi cross-coupling conditions (e.g., in the presence of a palladium catalyst). Cross-coupling reaction under coupling conditions (e.g., in the presence of a palladium catalyst) or trifluoromethylation conditions (e.g., in the presence of a copper catalyst) gives 1-13 . 1-13 and the adduct of Formula 1-14 (where M is boronic acid, boronic acid ester, or an appropriately substituted metal [e.g., M is B(OR) ₂ , Sn(Alkyl) _3, or Zn- Hal]), either standard Suzuki cross-coupling conditions (e.g., in the presence of a palladium catalyst and an appropriate base), or standard Stille cross-coupling conditions (e.g., in the presence of a palladium catalyst), or Intermediates 1-15 can be prepared by cross-coupling reaction under standard Negishi cross-coupling conditions (e.g., in the presence of a palladium catalyst). Intermediate 1-15 can be converted to intermediate 1-16 through oxidation of astragalus using a suitable oxidizing agent (e.g., m-CPBA) followed by SnAr reaction, or cross-coupling reaction (Org. Lett. 2002 , 4, 979-981). Removal of the protecting groups in 1-16 provides amine 1-17 . Subsequent functionalization of the resulting amine (e.g., coupling with an acid chloride, e.g., acryloyl chloride) provides the desired products 1-18 . The sequence of chemical reactions described above may be rearranged or omitted as appropriate to suit the preparation of different analogs.

KRAS protein

The Ras family includes three members: KRAS, NRAS, and HRAS. RAS mutant cancers account for approximately 25% of human cancers. KRAS is the most frequently mutated isoform in human cancers: 85% of all RAS mutations are KRAS, 12% are NRAS, and 3% are HRAS (Simanshu, D. et al. Cell 170.1 (2017):17- 33). KRAS mutations are prevalent among the top three most lethal cancer types: pancreatic cancer (97%), colorectal cancer (44%), and lung cancer (30%) (Cox, A.D. et al. Nat Rev Drug Discov (2014) 13: 828-51). Most RAS mutations occur at amino acid residues/codons 12, 13, and 61; Codon 12 mutations are the most frequent in KRAS. The frequency of specific mutations varies between RAS gene isoforms, but G12D mutations are most prevalent in KRAS, whereas Q61R and G12R mutations are most frequent in NRAS and HRAS. Additionally, the spectrum of mutations within RAS isoforms differs between cancer types. For example, KRAS G12D mutations are prevalent in pancreatic cancer (51%), followed by colorectal adenocarcinoma (45%) and lung cancer (17%) (Cox, A.D. et al. Nat Rev Drug Discov (2014) 13:828- 51). In contrast, KRAS G12C mutations are predominant in non-small cell lung cancer (NSCLC), comprising 11 to 16% of lung adenocarcinomas (nearly half of mutant KRAS are G12C), as well as 2 to 5% of pancreatic and colorectal adenocarcinomas, respectively. do (Cox, A.D. et al. Nat. Rev. Drug Discov. (2014) 13:828-51). Using shRNA knockdown of thousands of genes across hundreds of cancer cell lines, genomic studies have demonstrated that cancer cells expressing KRAS mutations are highly dependent on KRAS function for cell growth (McDonald, R. et al. Cell 170 (2017): 577-592). Taken together, these findings suggested that KRAS mutations play an important role in human cancer and that the development of inhibitors targeting mutant KRAS may be useful in the clinical treatment of diseases characterized by KRAS mutations.

How to use

Cancer types in which KRAS carrying G12C, G12V and G12D mutations are implicated include carcinoma (e.g., pancreatic, colorectal, lung, bladder, stomach, esophagus, breast, head and neck, cervical skin, thyroid); Hematopoietic malignancies (e.g., myeloproliferative neoplasms (MPNs), myelodysplastic syndromes (MDS), chronic and juvenile myelomonocytic leukemias (CMML and JMML), acute myeloid leukemia (AML), and acute lymphoblastic leukemias. (ALL) and multiple myeloma (MM)); and other neoplasms (e.g., glioblastoma and sarcoma). Additionally, KRAS mutations have been found to cause acquired resistance to anti-EGFR therapy (Knickelbein, K. et al. Genes & Cancer, (2015): 4-12). KRAS mutations can cause immunological and inflammatory disorders (Fernandez-Medarde, A. et al. Genes & Cancer, (2011): 344-358), such as Ras-associated lymphoproliferative disorder caused by somatic mutations in KRAS or NRAS. (RALD) or juvenile myelomonocytic leukemia (JMML).

Compounds of the present disclosure can inhibit the activity of KRAS protein. For example, the compounds of the present disclosure can be used to inhibit the activity of KRAS in a cell or individual or patient in need of inhibition of the enzyme by administering an inhibitory amount of one or more compounds of the present disclosure to the cell, individual, or patient. You can.

As KRAS inhibitors, the compounds of the present disclosure are useful in the treatment of various diseases associated with abnormal expression or activity of KRAS. Compounds that inhibit KRAS would be useful by inhibiting angiogenesis or providing a means to prevent growth or induce apoptosis in tumors. Accordingly, it is expected that the compounds of the present disclosure will prove useful in the treatment or prevention of proliferative disorders, such as cancer. In particular, tumors with activating mutations in receptor tyrosine kinases or upregulation of receptor tyrosine kinases may be particularly sensitive to inhibitors.

In one aspect, provided herein is a method of inhibiting KRAS activity, comprising contacting a compound of the disclosure with KRAS. In one embodiment, the contacting step includes administering the compound to the patient.

In one aspect, provided herein is a method of inhibiting a KRAS protein carrying a G12C mutation, comprising contacting a compound of the disclosure with KRAS.

In one aspect, provided herein is a method of inhibiting a KRAS protein carrying a G12D mutation, the method comprising contacting KRAS with a compound of the disclosure.

In one aspect, provided herein is a method of inhibiting a KRAS protein carrying a G12V mutation, the method comprising contacting KRAS with a compound of the disclosure.

In another aspect, provided herein is a method of treating a disease or disorder associated with inhibition of KRAS interaction, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any of the formulas disclosed herein or It includes administering a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method of treating a disease or disorder associated with inhibiting a KRAS protein carrying a G12C mutation, comprising administering to a patient in need thereof a therapeutically effective amount of the formulas disclosed herein. and administering a compound of any of the above or a pharmaceutically acceptable salt thereof.

In yet another aspect, provided herein is a method of treating cancer in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound disclosed herein, wherein Cancer is characterized by interaction with the KRAS protein carrying the G12C mutation.

In another aspect, provided herein is a method of treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound of any of the formulas disclosed herein, or a pharmaceutically acceptable salt thereof. Includes steps.

In one aspect, provided herein are methods for treating a disease or disorder associated with inhibition of a KRAS interaction or mutant thereof in a patient in need thereof, wherein the method is provided to the patient. administering a disclosed compound, or a pharmaceutically acceptable salt thereof, or a composition comprising a compound disclosed herein, or a pharmaceutically acceptable salt thereof, with another therapy or therapeutic agent as described herein.

In one embodiment, the cancer is selected from hematological cancer, sarcoma, lung cancer, gastrointestinal cancer, urogenital cancer, liver cancer, bone cancer, nervous system cancer, gynecological cancer, and skin cancer.

In another embodiment, the lung cancer is non-small cell lung cancer (NSCLC), small cell lung cancer, bronchogenic carcinoma, squamous cell bronchiogenic carcinoma, undifferentiated small cell bronchiogenic carcinoma, undifferentiated large cell bronchiogenic carcinoma, adenocarcinoma, bronchiogenic carcinoma, alveolar carcinoma, It is selected from bronchiolar carcinoma, bronchial adenoma, chondromatous hamartoma, mesothelioma, parous cell and non-parous cell carcinoma, bronchial adenoma and pleuropulmonary blastoma.

In another embodiment, the lung cancer is non-small cell lung cancer (NSCLC). In yet another embodiment, the lung cancer is adenocarcinoma.

In one embodiment, the gastrointestinal cancer is esophageal squamous cell carcinoma, esophageal adenocarcinoma, esophageal leiomyosarcoma, esophageal lymphoma, gastric carcinoma, gastric lymphoma, gastric leiomyosarcoma, exocrine pancreatic carcinoma, pancreatic ductal adenocarcinoma, pancreatic insulinoma, pancreatic glucagonoma, pancreatic Gastrinoma, pancreatic carcinoid, pancreatic vipoma, small intestine adenocarcinoma, small intestine lymphoma, small intestine carcinoid tumor, Kaposi's sarcoma, small intestine leiomyoma, small intestine hemangioma, small intestine lipoma, small intestine neurofibroma, small intestine fibroma, colon adenocarcinoma, colon tube It is selected from adenoma, colonic trophoblastic adenoma, colonic hamartoma, colonic leiomyoma, colorectal cancer, gallbladder cancer, and anal cancer.

In one embodiment, the gastrointestinal cancer is colorectal cancer.

In another embodiment, the cancer is carcinoma. In another embodiment, the carcinoma is selected from pancreatic carcinoma, colorectal carcinoma, lung carcinoma, bladder carcinoma, gastric carcinoma, esophageal carcinoma, breast carcinoma, head and neck carcinoma, cervical skin carcinoma, and thyroid carcinoma.

In yet another embodiment, the cancer is a hematopoietic malignancy. In one embodiment, the hematopoietic malignancy is selected from multiple myeloma, acute myeloid leukemia, and myeloproliferative neoplasm.

In another embodiment, the cancer is a neoplasm. In another embodiment, the neoplasm is glioblastoma or sarcoma.

In certain embodiments, the present disclosure provides methods of treating a KRAS-mediated disorder in a patient in need thereof, comprising administering to said patient a compound according to the invention or a pharmaceutically acceptable dose thereof. and administering the composition.

In some embodiments, diseases and indications treatable using the compounds of the present disclosure include, but are not limited to, hematological cancers, sarcomas, lung cancers, gastrointestinal cancers, genitourinary cancers, liver cancers, bone cancers, nervous system cancers, gynecological cancers, and skin cancers. Not limited.

Exemplary blood cancers include lymphomas and leukemias, such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML). , diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma, non-Hodgkin's lymphoma (including relapsed or refractory NHL and relapsed follicular), Hodgkin's lymphoma, myeloproliferative diseases (e.g., primary myelofibrosis ( PMF), polycythemia vera (PV), essential thrombocytosis (ET), 8p11 myeloproliferative syndrome, myelodysplastic syndrome (MDS), T-cell acute lymphoblastic lymphoma (T-ALL), These include multiple myeloma, cutaneous T-cell lymphoma, adult T-cell leukemia, Waldenström's macroglobulinemia, hairy cell lymphoma, marginal zone lymphoma, chronic myeloid lymphoma, and Burkitt's lymphoma.

Exemplary sarcomas include chondrosarcoma, Ewing's sarcoma, osteosarcoma, rhabdomyosarcoma, angiosarcoma, fibrosarcoma, liposarcoma, myxoma, rhabdomyoma, rhabdosarcoma, fibroma, lipoma, hamartoma, lymphosarcoma, leiomyosarcoma. and teratoma.

Exemplary lung cancers include non-small cell lung cancer (NSCLC), small cell lung cancer, bronchiogenic carcinoma, squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma, alveolar (bronchiolar) carcinoma, bronchial adenoma, chondromatous hamartoma, mesothelioma, parmicellar and non-celluloma. -Includes pavicellular and non-pavicellular carcinoma, bronchial adenoma, and pleuropulmonary blastoma.

Exemplary gastrointestinal cancers include esophageal cancer (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), gastric cancer (carcinoma, lymphoma, leiomyosarcoma), pancreatic cancer (exocrine pancreatic carcinoma, pancreatic adenocarcinoma, insulinoma, glucagonoma, gastrinoma, Carsi). noid tumor, vipoma), small intestine cancer (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), colon cancer (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma) , leiomyoma), colorectal cancer, gallbladder cancer, and anal cancer.

Exemplary genitourinary tract cancers include kidney cancer (adenocarcinoma, Wilms tumor [nephroblastoma], renal cell carcinoma), bladder and urethral cancer (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate cancer (adenocarcinoma, sarcoma), and testicular cancer ( Seminoma, teratoma, embryonal carcinoma, teratoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma) and urothelial carcinoma.

Exemplary liver cancers include hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, and hemangioma.

Exemplary bone cancers include, for example, osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticular cell sarcoma), multiple myeloma, malignant giant cell tumor chordoma, osseous Includes chordoma (osteocartilaginous exostoses), benign enchondroma, chondroblastoma, chondromyxofibroma, osteoid and giant cell tumor.

Exemplary nervous system cancers include skull cancer (osteoma, hemangioma, granuloma, xanthoma, osteitis deforming), meningeal cancer (meningioma, meningiosarcoma, glioma), brain cancer (astrocytoma, medulloblastoma, glioma, ependymoma, Germ cell tumor (pineal body tumor), glioblastoma, glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumor, neuro-ectodermal tumor), and spinal cord cancer (neurofibroma, meningioma, glioma, sarcoma), neuroblastoma, Includes Lhermitte-Duclos disease and pineal tumor.

Exemplary gynecologic cancers include breast (ductal carcinoma, lobular carcinoma, breast sarcoma, triple-negative breast cancer, HER2-positive breast cancer, inflammatory breast cancer, papillary carcinoma), uterine cancer (endometrial carcinoma), and cervical cancer (cervical carcinoma, preneoplastic carcinoma). Cervical dysplasia), ovarian cancer (ovarian carcinoma (serous cystic adenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa cell tumor, Serotoli-Leydig cell tumor, undifferentiated germ cell tumor, malignant teratoma), vulvar cancer (squamous) Includes epithelial cell carcinoma, carcinoma in situ, adenocarcinoma, fibrosarcoma, and melanoma), vaginal cancer (clear cell carcinoma, squamous cell carcinoma, staphylococcus sarcoma (embryonic rhabdomyosarcoma), and fallopian tube cancer (carcinoma).

Exemplary skin cancers include melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, Merkel cell skin cancer, dysplastic nevus, lipoma, hemangioma, dermatofibroma, and keloid.

Exemplary head and neck cancers include glioblastoma, melanoma, rhabdomyosarcoma, lymphosarcoma, osteosarcoma, squamous cell carcinoma, adenocarcinoma, oral cavity cancer, laryngeal cancer, nasopharyngeal cancer, nasal cavity and paranasal cavity, thyroid and parathyroid cancer, tumors of the eye, lip tumors, and squamous head. Includes squamous head and neck cancer.

Compounds of the present disclosure may also be useful for inhibiting tumor metastasis.

In addition to tumorigenic neoplasms, the compounds of the present invention may also be used to treat achondroplasia, hypochondrosis, dwarfism, thanatophoric dysplasia (TD) (clinical forms TD I and TD II), Apert syndrome, and crouzon. Including, but not limited to, Crouzon syndrome, Jackson-Weiss syndrome, Beare-Stevenson cutis gyrate syndrome, Pfeiffer syndrome, and craniosynostosis syndrome. It is useful in the treatment of skeletal and chondrocyte disorders. In some embodiments, the present disclosure provides methods for treating patients suffering from skeletal and chondrocyte disorders.

In some embodiments, the compounds described herein can be used to treat Alzheimer's disease, HIV, or tuberculosis.

As used herein, the term “8p11 myeloproliferative syndrome” is meant to refer to a myeloid/lymphoid neoplasm associated with abnormalities in FGFR1 and eosinophilia.

As used herein, the term “cell” is meant to refer to a cell in vitro, ex vivo, or in vivo. In some embodiments, ex vivo cells can be a portion of a tissue sample excised from an organism, such as a mammal. In some embodiments, in vitro cells can be cells in cell culture. In some embodiments, the in vivo cell is a living cell in an organism, such as a mammal.

As used herein, the term “contacting” refers to bringing together indicated moieties in an in vitro or in vivo system. For example, “contacting” KRAS with a compound described herein includes administration of a compound described herein to an individual or patient, such as a human having KRAS, as well as a sample containing cells or purified preparations containing KRAS. Including introducing a compound described herein into.

As used herein, the terms “individual”, “subject” or “patient”, used interchangeably, refer to mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, refers to any animal including horses or primates and most preferably humans.

As used herein, the phrase “therapeutically effective amount” refers to a substance disclosed herein that causes a biological or medical response in a tissue, system, animal, individual, or human to be sought by a researcher, veterinarian, physician, or other clinician. refers to the amount of an active compound or pharmaceutical agent, such as in any solid form or salt thereof. The appropriate “effective” amount in any individual case can be determined using techniques known to those skilled in the art.

The phrase "pharmaceutically acceptable" means that, within the scope of sound medical judgment, it can be used with human and animal tissues, without undue toxicity, irritation, allergic reaction, immunogenicity or other problems or complications, and commensurate with a reasonable benefit/risk ratio. It is used herein to refer to a compound, material, composition and/or dosage form suitable for use in contact.

As used herein, the phrase “pharmaceutically acceptable carrier or excipient” refers to a pharmaceutically acceptable substance, composition or vehicle, such as a liquid or solid filler, diluent, solvent or encapsulating material. Excipients or carriers are generally safe, non-toxic, not biologically or otherwise undesirable, and include excipients or carriers that are acceptable for human pharmaceutical use as well as veterinary use. In one embodiment, each ingredient is “pharmaceutically acceptable” as defined herein. See, e.g., Remington: The Science and Practice of Pharmacy, 21st ed.; Lippincott Williams & Wilkins: Philadelphia, Pa., 2005; Handbook of Pharmaceutical Excipients, 6th ed.; Rowe et al., Eds.; The Pharmaceutical Press and the American Pharmaceutical Association: 2009; Handbook of Pharmaceutical Additives, 3rd ed.; Ash and Ash Eds.; Gower Publishing Company: 2007; Pharmaceutical Preformulation and Formulation, 2nd ed.; Gibson Ed.; CRC Press LLC: Boca Raton, Fla., 2009].

As used herein, the terms “treating” or “treatment” include inhibiting a disease; For example, inhibiting a disease, condition or disorder (i.e., preventing further development of the pathology and/or symptoms) in an individual experiencing or exhibiting the pathology or symptoms of the disease, condition or disorder, or improving the disease. to do something; For example, improving a disease, condition or disorder (i.e. reversing the pathology and/or symptoms) in an individual experiencing or exhibiting the pathology or symptoms of the disease, condition or disorder, e.g., reducing the severity of the disease. It refers to what is ordered.

As used herein, the terms “prevent,” “preventing,” or “prevention” include prevention of at least one symptom associated with or caused by the condition, disease, or disorder being prevented.

For clarity, certain features of the invention that are described in the context of separate embodiments may also be provided in combination in a single embodiment (while such embodiments are intended to be combined as if written in multiple dependent forms). It is additionally recognized as Conversely, for the sake of brevity, various features of the invention that are described in the context of a single embodiment may also be provided separately or in any suitable subcombination.

combination therapy

I. Cancer therapy

Cancer cell growth and survival can be affected by dysfunction in multiple signaling pathways. Therefore, it is useful to combine different enzyme/protein/receptor inhibitors that modulate the activity of these conditions and represent different priorities of targets for treating these conditions. Targeting more than one signaling pathway (or more than one biological molecule implicated in a given signaling pathway) can reduce the likelihood of drug-resistance occurring in a cell population and/or reduce treatment toxicity.

One or more additional pharmaceutical agents, such as chemotherapy agents, anti-inflammatory agents, steroids, immunosuppressants, immuno-oncology agents, metabolic enzyme inhibitors, chemokine receptor inhibitors, and phosphatase inhibitors, as well as targeted therapies, such as , Bcr-Abl, Flt-3, EGFR, HER2, JAK, c-MET, VEGFR, PDGFR, c-Kit, IGF-1R, RAF, FAK and CDK4/6 kinase inhibitors, e.g. in WO 2006/056399 Those described can be used with the compounds of the present disclosure for the treatment of CDK2-related diseases, disorders or conditions. Other agents, such as therapeutic antibodies, can be used with the compounds of the disclosure for the treatment of CDK2-related diseases, disorders or conditions. One or more additional pharmaceutical agents may be administered to the patient simultaneously or sequentially.

In some embodiments, a CDK2 inhibitor can be administered or used in combination with a BCL2 inhibitor or a CDK4/6 inhibitor.

Compounds as disclosed herein can be used in combination with one or more other enzyme/protein/receptor inhibitor therapies for the treatment of diseases, such as cancer and other diseases or disorders described herein. Examples of diseases and indications treatable with combination therapy include those described herein. Examples of cancer include solid tumors and non-solid tumors such as liquid tumors, hematological cancers. Examples of infections include viral infections, bacterial infections, fungal infections, or parasitic infections. For example, the compounds of the present disclosure can be combined with one or more inhibitors of the following kinases for the treatment of cancer: Akt1, Akt2, Akt3, BCL2, CDK4/6, TGF-βR, PKA, PKG, PKC. , CaM-kinase, phosphorylase kinase, MEKK, ERK, MAPK, mTOR, EGFR, HER2, HER3, HER4, INS-R, IDH2, IGF-1R, IR-R, PDGFαR, PDGFβR, PI3K (alpha, beta , gamma, delta and multiple or selective), CSF1R, KIT, FLK-II, KDR/FLK-1, FLK-4, flt-1, FGFR1, FGFR2, FGFR3, FGFR4, c-Met, PARP, Ron, Sea, TRKA, TRKB, TRKC, TAM Kinase (Axl, Mer, Tyro3), FLT3, VEGFR/Flt2, Flt4, EphA1, EphA2, EphA3, EphB2, EphB4, Tie2, Src, Fyn, Lck, Fgr, Btk, Fak, SYK, FRK, JAK, ABL, ALK and B-Raf. In some embodiments, compounds of the present disclosure may be combined with one or more of the following inhibitors for the treatment of cancer or infection. Non-limiting examples of inhibitors that can be combined with the compounds of the present disclosure for the treatment of cancer and infection include FGFR inhibitors (FGFR1, FGFR2, FGFR3 or FGFR4, e.g., pemigatinib (INCB54828), INCB62079), EGFR inhibitors ( Also known as ErB-1 or HER-1; e.g., erlotinib, gefitinib, vandetanib, orsimertinib, cetuximab, necitumumab ( necitumumab or panitumumab), VEGFR inhibitors or pathway blockers (e.g., bevacizumab, pazopanib, sunitinib, sorafenib, axitinib) axitinib, regorafenib, ponatinib, cabozantinib, vandetanib, ramucirumab, lenvatinib, ziv-aflibercept )), PARP inhibitors (e.g. olaparib, rucaparib, veliparib or niraparib), JAK inhibitors (JAK1 and/or JAK2, e.g. ruxolitinib or baricitinib ; or JAK1 ; For example, itacitinib (INCB39110), INCB052793 or INCB054707 ), IDO inhibitors (e.g., epacadostat, NLG919 or BMS-986205, MK7162), LSD1 inhibitors (e.g., GSK2979552, INCB59872 and INCB60003), TDO inhibitors, PI3K-delta inhibitors (e.g., Parsacli parsaclisib (INCB50465) or INCB50797), PI3K-gamma inhibitors such as PI3K-gamma selective inhibitors, Pim inhibitors (e.g. INCB53914), CSF1R inhibitors, TAM receptor tyrosine kinases (Tyro-3, Axl and Mer; e.g. INCB081776), adenosine receptor antagonists (e.g. A2a/A2b receptor antagonists), HPK1 inhibitors, chemokine receptor inhibitors (e.g. CCR2 or CCR5 inhibitors), SHP1/2 phosphatase inhibitors, histone deacetylase inhibitors (HDAC), e.g. HDAC8 inhibitors , angiogenesis inhibitors, interleukin receptor inhibitors, bromo and extra terminal family member inhibitors (e.g. bromodomain inhibitors or BET inhibitors such as INCB54329 and INCB57643), c-MET inhibitors (e.g. capmatinib) ), anti-CD19 antibodies (e.g., tafasitamab), ALK2 inhibitors (e.g., INCB00928); or a combination thereof.

In some embodiments, a compound or salt described herein is administered in conjunction with a PI3Kδ inhibitor. In some embodiments, a compound or salt described herein is administered in combination with a JAK inhibitor. In some embodiments, a compound or salt described herein is administered in combination with a JAK1 or JAK2 inhibitor (e.g., baricitinib or ruxolitinib). In some embodiments, a compound or salt described herein is administered in combination with a JAK1 inhibitor. In some embodiments, a compound or salt described herein is administered with a JAK1 inhibitor that is selective over JAK2.

Additionally, for the treatment of cancer and other proliferative diseases, the compounds described herein may be useful in targeted therapies, such as c-MET inhibitors (e.g., capmatinib), anti-CD19 antibodies (e.g., tafasitamab), ALK2 inhibitor (eg, INCB00928); Or it can be used with a combination thereof.

Exemplary antibodies for use in combination therapy include trastuzumab (e.g., anti-HER2), ranibizumab (e.g., anti-VEGF-A), bevacizumab (AVASTIN™, e.g., anti-VEGF) ), panitumumab (e.g., anti-EGFR), cetuximab (e.g., anti-EGFR), rituxan (e.g., anti-CD20), and antibodies directed to c-MET, but these Not limited.

One or more of the following agents may be used in combination with the compounds of the present disclosure and are presented as a non-limiting list: cytostatic agent, cisplatin, doxorubicin, taxotere, taxol, etoposide, irinotecan, cam. camptosar, topotecan, paclitaxel, docetaxel, etotilone, tamoxifen, 5-fluorouracil, methotrexate, temozolomide, cyclophosphamide, SCH 66336, R115777, L778,123, BMS 214662, IRESSA™ (gefitinib), TARCEVA™ (erlotinib), antibodies against EGFR, intron, ara-C, adriamycin, cytoxan, gemcitabine, uracil mustard, chlormetine, ifosfamide, melphalan, chlora Busil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine , fludarabine phosphate, oxaliplatin, leucovirin, ELOXATIN™ (oxaliplatin), pentostatin, vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubi. Cin, mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, teniposide 17.alpha.-ethinylastradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, Dromostanolone propionate, testolactone, megestrol acetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianisene, hydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesterone acetate. , leuplolide flutamide, toremifene, goserelin, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane, mitoxathrone, levamisole, nabelbene, anastrazole, letrazole, Capecitabine, reloxapine, droloxapine, hexamethylmelamine, Avastin, HERCEPTIN™ (trastuzumab), BEXXAR™ (tositumomab), VELCADE™ (bortezomib), ZEVALIN™ (libritumomab tiuxetan) , TRISENOX™ (arsenic trioxide), , exemestane, ifosphamide, rituximab, C225 (cetuximab), Campath, (alemtuzumab), clofarabine, cladribine, apidicolone, rituxan, sunitinib , dasatinib, dezacitabine, Sml1, fludarabine, pentostatin, triapine, Dedox, Trimitox amidox, 3-AP and MDL-101,731.

The compounds of the present disclosure can further be used in combination with other methods of treating cancer, for example, by chemotherapy, irradiation therapy, tumor-targeted therapy, adjuvant therapy, immunotherapy or surgery. . Examples of immunotherapies include cytokine therapy (e.g., interferon, GM-CSF, G-CSF, IL-2), CRS-207 immunotherapy, cancer vaccines, monoclonal antibodies, bispecific or multi-specific antibodies. , antibody drug conjugates, adoptive T cell transfer, Toll receptor agonists, RIG-I agonists, oncolytic virotherapy, and immunomodulatory small molecules including thalidomide or JAK1/2 inhibitors and PI3Kδ inhibitors. The compound may be administered in combination with one or more anti-cancer drugs, such as chemotherapy agents. Examples of chemotherapy agents include any of the following: abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretinoin. Altretamine, anastrozole, arsenic trioxide, asparaginase, azacitidine, bevacizumab, bexarotene, baricitinib, bleomycin, bortezomib ( bortezomib), intravenous busulfan, oral busulfan, calusterone, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, Cladri Vin, clofarabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, dalteparin sodium, dasatinib, daunorubicin, decitabine, denileukin, denileukin deftitox, dex Razoxan, docetaxel, doxorubicin, drmostanolone propionate, eculizumab, epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, Floxuridine, fludarabine, fluorouracil, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin acetate, histrelin acetate, ibritumomab tiuxetan, idarubicin , ifosfamide, imatinib mesylate, interferon alpha 2a, irinotecan, lapatinib ditosylate, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole, lomustine, mechlorethamine, megestrol. Acetate, melphalan, mercaptopurine, methoxtrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone, nandrolone phenpropionate, nelarabine, nofetumomab, oxaliplatin, Paclitaxel, pamidronate, panitumumab, pegasparagase, pegfilgrastim, pemetrexed disodium, pentostatin, fifobroman, plicamycin, procarbazine, quinacrine, rasburicase ), rituximab, ruxolitinib, sorafenib, streptozocin, sunitinib, sunitinib maleate, tamoxifen, temozolomide, teniposide, testolactone, thalidomide, thioguanine, thiotepa, topotecan, toremi Fen, tositumomab, trastuzumab, tretinoin, uracil mustard, valubicin, vinblastine, vincristine, vinorelbine, vorinostat and zoledronate.

Additional examples of chemotherapy agents include proteasome inhibitors (e.g., bortezomib), thalidomide, revlimid, and DNA-damaging agents such as melphalan, doxorubicin, cyclophosphamide, vincristine, etoposide, camu. Including Steen et al.

Exemplary steroids include corticosteroids such as dexamethasone or prednisone.

Exemplary Bcr-Abl inhibitors include imatinib mesylate (GLEEVAC™), nilotinib, dasatinib, bosutinib, and ponatinib and pharmaceutically acceptable salts. Other exemplary suitable Bcr-Abl inhibitors include compounds of the genera and species disclosed in US Pat. No. 5,521,184, WO 04/005281, and US Serial No. 60/578,491, and pharmaceutically acceptable salts thereof.

Exemplary suitable Flt-3 inhibitors include midostaurin, lestaurtinib, linipanib, sunitinib, sunitinib, maleate, solatinib, quizartinib, creolanib, pacritinib, nandutinib, PLX3397 and ASP2215, and pharmaceutically acceptable salts thereof. Other exemplary suitable Flt-3 inhibitors include compounds and pharmaceutically acceptable salts thereof, such as those disclosed in WO 03/037347, WO 03/099771 and WO 04/046120.

Exemplary suitable RAF inhibitors include dabrafenib, sorafenib and vemurafenib and pharmaceutically acceptable salts thereof. Other exemplary suitable RAF inhibitors include compounds and pharmaceutically acceptable salts thereof, such as those disclosed in WO 00/09495 and WO 05/028444.

Exemplary suitable FAK inhibitors include VS-4718, VS-5095, VS-6062, VS-6063, BI853520 and GSK2256098, and pharmaceutically acceptable salts thereof. Other exemplary suitable FAK inhibitors include compounds and their pharmaceutically acceptable compounds, such as those disclosed in WO 04/080980, WO 04/056786, WO 03/024967, WO 01/064655, WO 00/053595 and WO 01/014402. Contains salt.

Examples of suitable CDK4/6 inhibitors include palbociclib, ribociclib, triraciclib lerosiclib and abemaciclib and their pharmaceutically acceptable salts. Examples of other suitable CDK4/6 inhibitors include compounds as disclosed in WO 09/085185, WO 12/129344, WO 11/101409, WO 03/062236, WO 10/075074, and WO 12/061156, and their Contains pharmaceutically acceptable salts.

In some embodiments, compounds of the present disclosure may be used in combination with one or more other kinase inhibitors, including imatinib, to treat patients who are resistant to imatinib or other kinase inhibitors.

In some embodiments, compounds of the present disclosure can be used in combination with chemotherapy agents in the treatment of cancer and can improve treatment response compared to the response to the chemotherapy agent alone, without exacerbating its toxic effects. In some embodiments, compounds of the present disclosure can be used in combination with chemotherapy provided herein. For example, additional pharmaceutical agents used in the treatment of multiple myeloma may include, without limitation, melphalan, melphalan + prednisone [MP], doxorubicin, dexamethasone, and Velcade (bortezomib). Additional additional agents used in the treatment of multiple myeloma include Bcr-Abl, Flt-3, RAF and FAK kinase inhibitors. In some embodiments, the agent is an alkylating agent, proteasome inhibitor, corticosteroid, or immunomodulatory agent. Examples of alkylating agents include cyclophosphamide (CY), melphalan (MEL), and bendamustine. In some embodiments, the proteasome inhibitor is carfilzomib. In some embodiments, the corticosteroid is dexamethasone (DEX). In some embodiments, the immunomodulatory agent is lenalidomide (LEN) or pomalidomide (POM). Additive or synergistic effects are desirable outcomes of combining CDK2 inhibitors of the present disclosure with additional agents.

These agents may be combined with the compounds of the invention in single or sequential dosage forms, or these agents may be administered simultaneously or sequentially as separate dosage forms.

Compounds of the present disclosure may be used in combination with one or more other inhibitors or one or more therapies for the treatment of infections. Examples of infections include viral infections, bacterial infections, fungal infections, or parasitic infections.

In some embodiments, a corticosteroid, such as dexamethasone, is administered to a patient in combination with a compound of the disclosure when dexamethasone is administered intermittently as opposed to continuously.

Compounds of formula (I) as described herein or any one of formulas, compounds as recited in any of the claims and as described herein, or salts thereof, can be used in the treatment of other immunogenic agents, such as cancerous cells, purified It can be used in combination with cells transfected with genes encoding tumor antigens (including recombinant proteins, peptides and carbohydrate molecules), cells, and immune stimulating cytokines. Non-limiting examples of tumor vaccines that can be used include peptides of melanoma antigens, such as gp100, MAGE antigen, Trp-2, MARTI and/or tyrosinase, or tumor cells transfected to express the cytokine GM-CSF. Includes.

A compound of formula (I) or any of the formulas as described herein, a compound as recited in any of the claims and as described herein, or salts thereof, can be used in combination with a vaccination protocol for the treatment of cancer. can be used In some embodiments, tumor cells are transduced to express GM-CSF. In some embodiments, tumor vaccines include proteins from viruses implicated in human cancer, such as human papillomavirus (HPV), hepatitis viruses (HBV and HCV), and Kaposi's herpes sarcoma virus (KHSV). In some embodiments, compounds of the present disclosure may be used in combination with tumor-specific antigens, such as heat shock, isolated from the tumor tissue itself. In some embodiments, a compound of formula (I) as described herein or any one of formulas, a compound as recited in any of the claims and as described herein, or salts thereof, produces a potent anti-tumor response. It can be combined with dendritic cell immunization to activate.

Compounds of the present disclosure can be used in combination with bispecific cytomegalopeptides that target Fe alpha or Fe gamma receptor-expressing effector cells to tumor cells. Compounds of the present disclosure can also be combined with cytomegalopeptides that activate host immune responses.

In some additional embodiments, combinations of compounds of the present disclosure with other therapeutic agents may be administered to patients before, during, and/or after bone marrow transplantation or stem cell transplantation. The compounds of the present disclosure can be used in combination with bone marrow transplantation for the treatment of various tumors of hematopoietic origin.

Compounds of formula (I) as described herein or any one of formulas, compounds as recited in any of the claims and as described herein, or salts thereof, induce an immune response to pathogens, toxins, and self-antigens. It can be used in combination with a vaccine to stimulate it. Examples of pathogens for which this therapeutic approach may be particularly useful include pathogens for which there are currently no effective vaccines, or pathogens for which conventional vaccines are less than fully effective. These are: HIV, hepatitis (types A, B and C), influenza, herpes, Giardia , malaria, Leishmania, Staphylococcus aureus and Pseudomonas aeruginosa . ), but is not limited to these.

Viruses that cause infections treatable by the methods of the present disclosure include human papillomavirus, influenza, hepatitis A, B, C or D virus, adenovirus, poxvirus, herpes simplex virus, human cytomegalovirus, severe acute respiratory infection. Syndromal viruses, Ebola virus, measles virus, herpes virus (e.g., VZV, HSV-1, HAV-6, HSV-II, and CMV, Epstein-Barr virus), flavivirus, echovirus, rhinovirus, coxsackie virus, Cornovirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papilloma virus, molluscum virus, poliovirus, rabies virus, JC virus. and arboviral encephalitis virus.

Pathogenic bacteria that cause infections treatable by the methods of the present disclosure include Chlamydia, Rickettsia bacteria, Mycobacteria, Staphylococcus, Streptococcus, Pneumococcus, Meningococci and Neisseria gonorrhoeae, Klebsiella, Proteus, Cellatia, Pseudomonas, Includes, but is not limited to, Legionella, diphtheria, salmonella, bacillus, cholera, tetanus, botulism, anthrax, plague, leptospirosis, and Lyme disease bacteria.

Pathogenic parasites that cause infections treatable by the methods of the present disclosure include Candida ( albicans , krusei , glabrata , tropicalis , etc.), Cryptococcus Cryptococcus neoformans , Aspergillus ( fumigatus , niger , etc.), Genus Mucorales ( mucor ), absidia , ), Rhizophus ), Sporothrix schenkii , Blastomyces dermatitidis, Paracoccidioides brasiliensis , Coccidioides imitis ( Coccidioides immitis ) and Histoplasma capsulatum , but are not limited to these.

Pathogenic parasites that cause infections treatable by the methods of the present disclosure include Shigella amoeba, Col. balantii, Fowler's amoeba, Acanthamoeba spp., Giardia lambia , Cryptosporidium sp. , Pneumocystis carinii , Plasmodium vivax, Babesia microti , Trypanosoma brucei , Trypanosoma cruzi , Leishmania donovani ( Leishmania donovani ), Toxoplasma gondi and Nippostrongylus brasiliensis .

When more than one pharmaceutical agent is administered to a patient, they may be administered simultaneously (e.g., in the case of more than two agents), individually, sequentially, or in combination.

Safe and effective methods of administering most of these chemotherapeutic agents are known to those skilled in the art. Additionally, such administration is described in standard literature. For example, multiple administrations of chemotherapy agents are described in the "Physicians' Desk Reference" (PDR, e.g., 1996 edition, Medical Economics Company, Montvale, NJ), the disclosure of which is as if set forth in its entirety. As if incorporated herein by reference.

II. immune checkpoint therapy

Compounds of the present disclosure can be used in combination with one or more immune checkpoint inhibitors for the treatment of diseases such as cancer or infections. Exemplary immune checkpoint inhibitors include inhibitors to immune checkpoint molecules, such as CBL-B, CD20, CD28, CD40, CD70, CD122, CD96, CD73, CD47, CDK2, GITR, CSF1R, JAK, PI3K delta, PI3K gamma, TAM , Araginase, HPK1, CD137 (also known as 4-1BB), ICOS, A2AR, B7-H3, B7-H4, BTLA, CTLA-4, LAG3, TIM3, TLR (TLR7/8), TIGIT, CD112R, VISTA , PD-1, PD-L1 and PD-L2. In some embodiments, the immune checkpoint molecule is a stimulatory checkpoint molecule selected from CD27, CD28, CD40, ICOS, OX40, GITR, and CD137. In some embodiments, the immune checkpoint molecule is an inhibitory checkpoint molecule selected from A2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3, PD-1, TIM3, TIGIT, and VISTA. In some embodiments, the compounds provided herein may be used in combination with one or more agents selected from KIR inhibitors, TIGIT inhibitors, LAIR1 inhibitors, CD160 inhibitors, 2B4 inhibitors, and TGFR beta inhibitors.

In some embodiments, the compounds provided herein may be used in combination with one or more agents of immune checkpoint molecules, such as OX40, CD27, GITR, and CD137 (also known as 4-1BB).

In some embodiments, the inhibitor of the immune checkpoint molecule is an anti-PD1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody.

In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of PD-1 or PD-L1, such as an anti-PD-1 or anti-PD-L1 monoclonal antibody. In some embodiments, the anti-PD-1 or anti-PD-L1 antibody is nivolumab, pembrolizumab, atezolizumab, durvalumab, avelumab, cemiplimab, atezolizumab, avelumab, tislelizumab. (tislelizumab), spartalizumab (PDR001), cetrelimab (JNJ-63723283), toripalimab (JS001), camrelizumab (SHR-1210), sintilimab (IBI308), AB122 (GLS-010) , AMP-224, AMP-514/MEDI-0680, BMS936559, JTX-4014, BGB-108, SHR-1210, MEDI4736, FAZ053, BCD-100, KN035, CS1001, BAT1306, LZM009, AK105, HLX10, SHR-1316 , CBT-502 (TQB2450), A167 (KL-A167), STI-A101 (ZKAB001), CK-301, BGB-A333, MSB-2311, HLX20, TSR-042 or LY3300054. In some embodiments, inhibitors of PD-1 or PD-L1 are disclosed in U.S. Patents 7,488,802, 7,943,743, 8,008,449, 8,168,757, 8,217,149, or 10,308,644; US Patent Publication Nos. 2017/0145025, 2017/0174671, 2017/0174679, 2017/0320875, 2017/0342060, 2017/0362253, 2018/0016260, 2018/0057486, 2018/01 77784, 2018/0177870, 2018/0179179, 2018/ 0179201, 2018/0179202, 2018/0273519, 2019/0040082, 2019/0062345, 2019/0071439, 2019/0127467, 2019/0144439, 2019/0202824, 20 19/0225601, 2019/0300524, or 2019/0345170; or PCT Publication No. WO 03042402, WO 2008156712, WO 2010089411, WO 2010036959, WO 2011066342, WO 2011159877, WO 2011082400, or WO 2011161699 (each of which is incorporated by reference in its entirety) incorporated herein by reference). In some embodiments, the inhibitor of PD-L1 is INCB086550.

In some embodiments, the antibody is an anti-PD-1 antibody, such as an anti-PD-1 monoclonal antibody. In some embodiments, the anti-PD-1 antibody is nivolumab, pembrolizumab, cemiplimab, spartalizumab, camrelizumab, cetrelimab, toripalimab, sintilimab, AB122, AMP. -224, JTX-4014, BGB-108, BCD-100, BAT1306, LZM009, AK105, HLX10 or TSR-042. In some embodiments, the anti-PD-1 antibody is nivolumab, pembrolizumab, cemiplimab, spartalizumab, camrelizumab, cetrelimab, toripalimab, or sintilimab. In some embodiments, the anti-PD-1 antibody is pembrolizumab. In some embodiments, the anti-PD-1 antibody is nivolumab. In some embodiments, the anti-PD-1 antibody is cemiplimab. In some embodiments, the anti-PD-1 antibody is spartalizumab. In some embodiments, the anti-PD-1 antibody is camrelizumab. In some embodiments, the anti-PD-1 antibody is cetrelimab. In some embodiments, the anti-PD-1 antibody is toripalimab. In some embodiments, the anti-PD-1 antibody is sintilimab. In some embodiments, the anti-PD-1 antibody is AB122. In some embodiments, the anti-PD-1 antibody is AMP-224. In some embodiments, the anti-PD-1 antibody is JTX-4014. In some embodiments, the anti-PD-1 antibody is BGB-108. In some embodiments, the anti-PD-1 antibody is BCD-100. In some embodiments, the anti-PD-1 antibody is BAT1306. In some embodiments, the anti-PD-1 antibody is LZM009. In some embodiments, the anti-PD-1 antibody is AK105. In some embodiments, the anti-PD-1 antibody is HLX10. In some embodiments, the anti-PD-1 antibody is TSR-042. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab or pembrolizumab. In some embodiments, the anti-PD-1 monoclonal antibody is MGA012 (INCMGA0012; retifanlimab). In some embodiments, the anti-PD1 antibody is SHR-1210. Other anti-cancer agent(s) include antibody therapies, such as 4-1BB (e.g., urelumumab, utomilumab). In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of PD-L1, such as an anti-PD-L1 monoclonal antibody. In some embodiments, the anti-PD-L1 monoclonal antibody is atezolizumab, avelumab, durvalumab, tislelizumab, BMS-935559, MEDI4736, atezolizumab (MPDL3280A; also known as RG7446), Avelumab (MSB0010718C), FAZ053, KN035, CS1001, SHR-1316, CBT-502, A167, STI-A101, CK-301, BGB-A333, MSB-2311, HLX20 or LY3300054. In some embodiments, the anti-PD-L1 antibody is atezolizumab, avelumab, durvalumab, or tislelizumab. In some embodiments, the anti-PD-L1 antibody is atezolizumab. In some embodiments, the anti-PD-L1 antibody is avelumab. In some embodiments, the anti-PD-L1 antibody is durvalumab. In some embodiments, the anti-PD-L1 antibody is tislelizumab. In some embodiments, the anti-PD-L1 antibody is BMS-935559. In some embodiments, the anti-PD-L1 antibody is MEDI4736. In some embodiments, the anti-PD-L1 antibody is FAZ053. In some embodiments, the anti-PD-L1 antibody is KN035. In some embodiments, the anti-PD-L1 antibody is CS1001. In some embodiments, the anti-PD-L1 antibody is SHR-1316. In some embodiments, the anti-PD-L1 antibody is CBT-502. In some embodiments, the anti-PD-L1 antibody is A167. In some embodiments, the anti-PD-L1 antibody is STI-A101. In some embodiments, the anti-PD-L1 antibody is CK-301. In some embodiments, the anti-PD-L1 antibody is BGB-A333. In some embodiments, the anti-PD-L1 antibody is MSB-2311. In some embodiments, the anti-PD-L1 antibody is HLX20. In some embodiments, the anti-PD-L1 antibody is LY3300054.

In some embodiments, the inhibitor of an immune checkpoint molecule is a small molecule that binds PD-L1 or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of an immune checkpoint molecule is a small molecule that binds to and is internalized by PD-L1 or a pharmaceutically acceptable salt thereof. In some embodiments, inhibitors of immune checkpoint molecules are those described in US 2018/0179201, US 2018/0179197, US 2018/0179179, US 2018/0179202, US 2018/0177784, US 2018/0177870, US Patent Application No. 16/369,654 ( Filed March 29, 2019) and U.S. Patent Application No. 62/688,164, or pharmaceutically acceptable salts thereof, each of which is incorporated herein by reference in its entirety.

In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of KIR, TIGIT, LAIR1, CD160, 2B4, and TGFR beta.

In some embodiments, the inhibitor is MCLA-145.

In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of CTLA-4, such as an anti-CTLA-4 antibody. In some embodiments, the anti-CTLA-4 antibody is ipilimumab, tremelimumab, AGEN1884, or CP-675,206.

In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of LAG3, such as an anti-LAG3 antibody. In some embodiments, the anti-LAG3 antibody is BMS-986016, LAG525, INCAGN2385, or efthylagimod alfa (IMP321).

In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of CD73. In some embodiments, the inhibitor of CD73 is oleclumab.

In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of TIGIT. In some embodiments, the inhibitor of TIGIT is OMP-31M32.

In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of VISTA. In some embodiments, the inhibitor of VISTA is JNJ-61610588 or CA-170.

In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of B7-H3. In some embodiments, the inhibitor of B7-H3 is enoblituzumab, MGD009, or 8H9.

In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of a KIR. In some embodiments, the inhibitor of KIR is lirilumab or IPH4102.

In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of A2aR. In some embodiments, the inhibitor of A2aR is CPI-444.

In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of TGF-beta. In some embodiments, the inhibitor of TGF-beta is trabedersen, galusertinib, or M7824.

In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of PI3K-gamma. In some embodiments, the inhibitor of PI3K-gamma is IPI-549.

In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of CD47. In some embodiments, the inhibitor of CD47 is Hu5F9-G4 or TTI-621.

In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of CD73. In some embodiments, the inhibitor of CD73 is MEDI9447.

In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of CD70. In some embodiments, the inhibitor of CD70 is cusatuzumab or BMS-936561.

In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of TIM3, such as an anti-TIM3 antibody. In some embodiments, the anti-TIM3 antibody is INCAGN2390, MBG453, or TSR-022.

In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of CD20, such as an anti-CD20 antibody. In some embodiments, the anti-CD20 antibody is obinutuzumab or rituximab.

In some embodiments, the agonist of an immune checkpoint molecule is an agonist of OX40, CD27, CD28, GITR, ICOS, CD40, TLR7/8, and CD137 (also known as 4-1BB).

In some embodiments, the agonist of CD137 is urelumab. In some embodiments, the agonist of CD137 is utomilumab.

In some embodiments, the agonist of an immune checkpoint molecule is an inhibitor of GITR. In some embodiments, the agonist of GITR is TRX518, MK-4166, INCAGN1876, MK-1248, AMG228, BMS-986156, GWN323, MEDI1873, or MEDI6469. In some embodiments, the agent of the immune checkpoint molecule is an agonist of OX40, such as an OX40 agonist antibody or OX40L fusion protein. In some embodiments, the anti-OX40 antibody is INCAGN01949, MEDI0562 (tabolimab), MOXR-0916, PF-04518600, GSK3174998, BMS-986178, or 9B12. In some embodiments, the OX40L fusion protein is MEDI6383.

In some embodiments, the agonist of an immune checkpoint molecule is an agonist of CD40. In some embodiments, the agonist of CD40 is CP-870893, ADC-1013, CDX-1140, SEA-CD40, RO7009789, JNJ-64457107, APX-005M, or Chi Lob 7/4.

In some embodiments, the agonist of an immune checkpoint molecule is an agonist of ICOS. In some embodiments, the agent of ICOS is GSK-3359609, JTX-2011, or MEDI-570.

In some embodiments, the agonist of the immune checkpoint molecule is an agonist of CD28. In some embodiments, the agonist of CD28 is theralizumab.

In some embodiments, the agonist of the immune checkpoint molecule is an agonist of CD27. In some embodiments, the agonist of CD27 is varlilumab.

In some embodiments, the agonist of an immune checkpoint molecule is an agonist of TLR7/8. In some embodiments, the agonist of TLR7/8 is MEDI9197.

Compounds of the present disclosure can be used in combination with bispecific antibodies. In some embodiments, one of the domains of the bispecific antibody targets PD-1, PD-L1, CTLA-4, GITR, OX40, TIM3, LAG3, CD137, ICOS, CD3, or TGFβ receptor. In some embodiments, the bispecific antibody binds PD-1 and PD-L1. In some embodiments, the bispecific antibody that binds PD-1 and PD-L1 is MCLA-136. In some embodiments, the bispecific antibody binds PD-L1 and CTLA-4. In some embodiments, the bispecific antibody that binds PD-L1 and CTLA-4 is AK104.

In some embodiments, compounds of the present disclosure may be used in combination with one or more metabolic enzyme inhibitors. In some embodiments, the metabolic enzyme inhibitor is an inhibitor of IDO1, TDO, or arginase. Examples of IDO1 inhibitors include epacadostat, NLG919, BMS-986205, PF-06840003, IOM2983, RG-70099, and LY338196. Arginase inhibitors include INCB1158.

As provided throughout, additional compounds, inhibitors, agents, etc. may be combined with the compounds of the invention in single or sequential dosage forms, or they may be administered simultaneously or sequentially as separate dosage forms.

Formulations, Dosage Forms and Administration

Compounds of the present disclosure, when used as pharmaceuticals, may be administered in the form of pharmaceutical compositions. Accordingly, the present disclosure refers to a compound of Formula I, II, or any of the formulas as described herein, a compound as recited in any of the claims and as described herein, or a pharmaceutically acceptable salt thereof, or A composition comprising any of the embodiments thereof and at least one pharmaceutically acceptable carrier or excipient is provided. These compositions can be prepared in a manner well known in the pharmaceutical arts and administered by a variety of routes depending on whether local or systemic treatment is indicated and on the area to be treated. Administration may be topical (including transdermal, epithelial, ocular, and mucosal, including intranasal, vaginal, and rectal delivery), pulmonary (e.g., by inhalation or aeration of powders or aerosols, including by nebulizer; intratracheal or intranasally), orally or parenterally. Parenteral administration may be administered intravenously, intraarterially, subcutaneously, intraperitoneally, intramuscularly, or by injection or infusion; or intracranial, such as intrathecal or intraventricular administration. Parenteral administration may be in the form of a single bolus dose or, for example, by a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners, etc. may be necessary or desirable.

The present invention also includes pharmaceutical compositions containing, as an active ingredient, a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, in combination with one or more pharmaceutically acceptable carriers or excipients. In some embodiments, the composition is suitable for topical administration. In preparing the compositions of the present invention, the active ingredients are typically mixed with, diluted with, or enclosed in such carriers, such as in the form of capsules, sachets, paper, or other containers. When an excipient acts as a diluent, it can be a solid, semi-solid or liquid substance that acts as a vehicle, carrier or medium for the active ingredient. Accordingly, the compositions can be tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), e.g., containing up to 10% by weight of the active compound. It may be in the form of ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaging powders containing .

In preparing formulations, the active compound may be milled to provide an appropriate particle size prior to combining with other ingredients. If the active compound is substantially insoluble, it may be ground to a particle size of less than 200 mesh. If the active compound is substantially water-soluble, the particle size can be adjusted by milling to provide a substantially uniform distribution in the formulation, e.g., about 40 mesh.

The compounds of the invention may be milled using known milling procedures, such as wet milling, to obtain appropriate particle sizes for tablet formation and for other dosage forms. Finely divided (nanoparticulate) preparations of the compounds of the invention can be prepared by methods known in the art, for example in WO 2002/000196.

Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, Contains water, syrup and methyl cellulose. The formulation may further include lubricants such as talc, magnesium stearate and mineral oil; humectant; emulsion and suspension formulations; Preservatives such as methyl- and propylhydroxy-benzoate; sweetener; and flavoring agents. Compositions of the invention can be formulated to provide rapid, sustained or delayed release of the active ingredient after administration to a patient using procedures known in the art.

In some embodiments, the pharmaceutical composition comprises silicified microcrystalline cellulose (SMCC) and at least one compound described herein, or a pharmaceutically acceptable salt thereof. In some embodiments, the silicified microcrystalline cellulose comprises about 98% microcrystalline cellulose and about 2% silicon dioxide (w/w).

In some embodiments, the composition is a sustained release composition comprising at least one compound described herein, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient. In some embodiments, the composition comprises at least one compound described herein, or a pharmaceutically acceptable salt thereof, and at least one component selected from microcrystalline cellulose, lactose monohydrate, hydroxypropyl methylcellulose, and polyethylene oxide. Includes. In some embodiments, the composition comprises at least one compound described herein, or a pharmaceutically acceptable salt thereof, and microcrystalline cellulose, lactose monohydrate, and hydroxypropyl methylcellulose. In some embodiments, the composition includes a salt described herein and microcrystalline cellulose, lactose monohydrate, and polyethylene oxide. In some embodiments, the composition further includes magnesium stearate or silicon dioxide. In some embodiments, the microcrystalline cellulose is Avicel PH102™. In some embodiments, the lactose monohydrate is Fast-flo 316™. In some embodiments, the hydroxypropyl methylcellulose is hydroxypropyl methylcellulose 2208 K4M (e.g., Methocel K4 M Premier™ and/or hydroxypropyl methylcellulose 2208 K100LV (e.g., Methocel K00LV™). In some embodiments, In this case, the polyethylene oxide is polyethylene oxide WSR 1105 (eg, Polyox WSR 1105™).

In some embodiments, wet granulation methods are used to produce the composition. In some embodiments, dry granulation methods are used to produce the composition.

The composition may be formulated in unit dosage form, with each dosage containing from about 5 to about 1,000 mg (1 g), more typically from about 100 mg to about 500 mg, of the active ingredient. In some embodiments, each dosage contains about 10 mg of active ingredient. In some embodiments, each dosage contains about 50 mg of active ingredient. In some embodiments, each dosage contains about 25 mg of active ingredient. The term “unit dosage form” refers to physically discrete units suitable for single dosage for human subjects and other mammals, each unit being combined with a suitable pharmaceutical excipient to produce the desired therapeutic effect. Contains a calculated, predetermined amount of active substance.

The ingredients used to formulate pharmaceutical compositions are of high purity and substantially free of potentially harmful contaminants (e.g., at least national food grade, usually at least analytical grade, and more typically at least pharmaceutical grade). ). Particularly for human consumption, the composition is preferably manufactured or formulated under good manufacturing practice standards as prescribed by applicable regulations of the U.S. Food and Drug Administration. For example, suitable formulations can be sterile and/or substantially isotonic and/or fully comply with all good manufacturing practice regulations of the U.S. Food and Drug Administration.

The active compounds can be effective over a wide dosage range and are generally administered in therapeutically effective amounts. However, the amount of compound actually administered will typically be determined by the medical staff depending on the relevant circumstances, including the condition being treated, the route of administration chosen, the actual compound administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, etc. It will be understood that

The therapeutic dosage of a compound of the invention may vary depending, for example, on the particular use for which treatment is to be effected, the mode of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of a compound of the invention in a pharmaceutical composition may vary depending on a number of factors, including dosage, chemical characteristics (e.g., hydrophobicity), and route of administration. For example, the compounds of the present invention can be provided as an aqueous physiological buffer solution containing about 0.1 to about 10% w/v of the compound for parenteral administration. Some typical dosage ranges are from about 1 g/kg to about 1 g/kg body weight per day. In some embodiments, the dosage range is from about 0.01 mg/kg to about 100 mg/kg body weight per day. Dosage will depend on variables such as the type and extent of the disease or disorder, the overall health of the particular patient, the relative biological potency of the selected compound, the formulation of the excipients, and the route of administration. Effective doses can be estimated from dose-response curves derived from in vitro or animal model test systems.

To prepare solid compositions, such as tablets, the principal active ingredients are mixed with pharmaceutical excipients to form a solid preformulation composition containing a homogeneous mixture of the compounds of the invention. When such preformulated compositions are referred to as homogeneous, the active ingredients are typically dispersed uniformly throughout the composition so that the composition can be easily subdivided into equally effective unit dosage forms such as tablets, pills, and capsules. These solid preformulations are then further subdivided into unit dosage forms of the type described above containing, for example, about 0.1 to about 1000 mg of the active ingredient of the invention.

Tablets or pills of the invention may be coated or otherwise compounded to provide dosage forms that provide the advantage of long-term action. For example, a tablet or pill may contain internally administered and externally administered components, the latter taking the form of an envelope over the former. The two components can be separated by an enteric layer, which serves to prevent decomposition in the stomach and allows the internal components to pass intact into the duodenum or be released with a delay. A variety of materials can be used for these enteric layers or coatings, including many polymeric acids and mixtures of polymeric acids and materials such as shellac, cetyl alcohol and cellulose acetate.

Liquid forms in which the compounds and compositions of the invention may be incorporated for oral or injectable administration include aqueous solutions, suitably flavored syrups, aqueous or oil suspensions, edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil. as well as flavored emulsions with elixirs and similar pharmaceutical vehicles.

Compositions for inhalation or inhalation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, and powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, the composition is administered by the oral or nasal route for local or systemic effects. The composition can be nebulized by use of an inert gas. The nebulized solution can be breathed directly from the nebulizing device, or the nebulizing device can be attached to a face mask, tent, or intermittent positive pressure breathing machine. Solution, suspension or powder compositions can be administered orally or nasally from a device that delivers the formulation in any suitable manner.

Topical formulations may contain one or more conventional carriers. In some embodiments, the ointment may contain water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white Vaseline, etc. The carrier composition of the cream may be based on water in combination with glycerol and one or more other ingredients such as glycerin monostearate, PEG-glycerin monostearate and cetylstearyl alcohol. Gels can be formulated using isopropyl alcohol and water, for example, in suitable combination with other ingredients such as glycerol, hydroxyethyl cellulose, etc. In some embodiments, the topical formulation contains at least about 0.1, at least about 0.25, at least about 0.5, at least about 1, at least about 2, or at least about 5% by weight of a compound of the invention. The topical formulation may be suitably packaged, e.g., in a tube of 100 g, optionally associated with instructions for the treatment of a selected indication, e.g., psoriasis or other skin condition.

The amount of compound or composition administered to a patient will vary depending on what is being administered, the purpose of administration, such as prevention or treatment, the patient's condition, the mode of administration, etc. In therapeutic applications, the composition may be administered to a patient already suffering from the disease in an amount sufficient to cure or at least partially arrest the symptoms of the disease and its complications. The effective dose will depend on the judgment of the attending clinician, depending on factors such as the severity of the disease, the age, weight, and general condition of the patient, as well as the disease condition being treated.

The composition administered to the patient may take the form of the pharmaceutical composition described above. These compositions may be sterilized by conventional sterilization techniques or may be sterile filtered. Aqueous solutions can be packaged for use as is, or can be lyophilized, and the lyophilized formulation is combined with a sterile aqueous carrier prior to administration. The pH of the compound formulation will typically be 3 to 11, more preferably 5 to 9, and most preferably 7 to 8. It will be understood that the use of any of the above excipients, carriers or stabilizers will result in the formation of pharmaceutical salts.

The therapeutic dosage of a compound of the invention may vary depending, for example, on the particular use for which treatment is to be effected, the mode of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of a compound of the invention in a pharmaceutical composition may vary depending on a number of factors, including dosage, chemical characteristics (e.g., hydrophobicity), and route of administration. For example, the compounds of the present invention can be provided as an aqueous physiological buffer solution containing about 0.1 to about 10% w/v of the compound for parenteral administration. Some typical dosage ranges are from about 1 □g/kg to about 1 g/kg body weight per day. In some embodiments, the dosage range is from about 0.01 mg/kg to about 100 mg/kg of body weight per day. Dosage will depend on variables such as the type and extent of the disease or disorder, the overall health of the particular patient, the relative biological potency of the selected compound, the formulation of the excipients, and the route of administration. Effective doses can be estimated from dose-response curves derived from in vitro or animal model test systems.

Labeled Compounds and Assay Methods

Another aspect of the invention is to localize and quantify KRAS proteins in tissue samples, including humans, and to identify KRAS ligands by inhibitory binding of labeled compounds, as well as in assays both in vitro and in vivo. as well as labeled compounds (radio-labeled, fluorescently labeled, etc.) of the present disclosure that would also be useful in imaging modalities. Substitution of one or more atoms of the compounds of the present disclosure may also be useful to result in differential ADME (adsorption, distribution, metabolism and excretion). Accordingly, the present invention provides a KRAS binding agent containing such labeled or substituted compounds. Includes assay methods.

The present disclosure further includes isotopically-labeled compounds of the disclosure. An “isotopically” or “radiolabeled” compound is one in which one or more atoms are replaced or substituted by an atom having an atomic mass or mass number that is different from the atomic mass or mass number typically found in nature (i.e., native). It is a compound of the disclosure. Suitable radionuclides that can be incorporated into the compounds of the present disclosure include ² H (also denoted as D for deuterium), ³ H (also denoted as T for tritium), ¹¹ C, ¹³ C, ¹⁴ C, Contains ¹³ N ^{, 15} N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ¹⁸ F, ³⁵ S, 36 Cl, ⁸² Br, ⁷⁵ Br, ⁷⁶ Br, ⁷⁷ Br, ¹²³ I, ¹²⁴ I, ¹²⁵ I and ¹³¹ I , but is not limited to these. For example, one or more hydrogen atoms in a compound of the present disclosure may be replaced with a deuterium atom (e.g., one or more hydrogen atoms of a C _1-6 alkyl group of Formulas I, II, or any of the formulas provided herein may optionally be replaced with a deuterium atom). may be replaced with a deuterium atom, for example, -CH ₃ is replaced with -CD ₃ ). In some embodiments, an alkyl group in Formula I, II, or any formula provided herein may be substituted with deuterium.

One or more member atoms of the compounds presented herein may be replaced or substituted with isotopes of the atom in natural or non-natural abundance. In some embodiments, the compound includes at least one deuterium. In some embodiments, the compound contains two or more deuterium atoms. In some embodiments, the compound contains 1 to 2, 1 to 3, 1 to 4, 1 to 5, or 1 to 6 deuterium atoms. In some embodiments, all of the hydrogen atoms in the compound may be replaced or substituted with deuterium atoms.

Synthetic methods for incorporating isotopes into organic compounds are known in the art (Deuterium Labeling in Organic Chemistry by Alan F. Thomas (New York, N.Y., Appleton-Century-Crofts, 1971; The Renaissance of H/D Exchange by Jens Atzrodt, Volker Derdau, Thorsten Fey and Jochen Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765; The Organic Chemistry of Isotopic Labeling by James R. Hanson, Royal Society of Chemistry, 2011). The obtained compounds can be used in various studies, such as NMR spectroscopy, metabolic experiments, and/or assays.

Substitution with heavier isotopes, such as deuterium, may offer certain therapeutic advantages due to greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and may therefore be desirable in some situations. (e.g., A. Kerekes et.al. J. Med. Chem. 2011, 54 , 201-210; R. Xu et.al. J. Label Compd. Radiopharm. 2015, 58 , 308-312) reference). In particular, substitutions at one or more metabolic sites may provide one or more therapeutic advantages.

The radionuclide incorporated into the radiolabeled compound of the invention will depend on the particular use of the radiolabeled compound. For example, for in vitro adenosine receptor labeling and competition assays, compounds incorporating ³ H, ¹⁴ C, ⁸² Br, ¹²⁵ I, ¹³¹ I, or ³⁵ S may be useful. For radio-imaging applications, ¹¹ C, ¹⁸ F, ¹²⁵ I, ¹²³ I, ¹²⁴ I, ¹³¹ I, ⁷⁵ Br, ⁷⁶ Br or ⁷⁷ Br may be useful.

It is understood that a “radio-labeled” or “labeled compound” is a compound that has incorporated at least one radionuclide. In some embodiments, the radionuclide is selected from ³ H, ¹⁴ C, ¹²⁵ I, ³⁵ S, and ⁸² Br.

The present disclosure may further include synthetic methods for incorporating radio-isotopes into the compounds of the disclosure. Synthetic methods for incorporating radio-isotopes into organic compounds are well known in the art, and those skilled in the art will readily recognize methods applicable to the compounds of the present disclosure.

Labeled compounds of the invention can be used in screening assays to identify and/or evaluate compounds. For example, a newly synthesized or identified compound that is labeled (i.e., a test compound) can be tested for its ability to bind to and activate the KRAS protein when it comes into contact with KRAS, by tracking the label and monitoring changes in its concentration. can be evaluated. For example, a test compound (labeled) can be evaluated for its ability to reduce the binding of another compound known to bind to KRAS (i.e., a standard compound). Therefore, the ability of a test compound to compete with a standard compound for binding to a KRAS protein is directly correlated to its binding affinity. Conversely, in some other screening assays, the standard compound is labeled and the test compound is unlabeled. Accordingly, the concentration of the labeled standard compound is monitored to assess competition between the standard compound and the test compound, thereby determining the relative binding affinity of the test compound.

kit

The present disclosure also relates to, for example, diseases or disorders associated with the activity of KRAS, comprising one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I), (II) or any of the embodiments thereof. For example, it includes pharmaceutical kits useful in the treatment or prevention of cancer or infection. Such kits may include one or more of a variety of conventional pharmaceutical kit components as will be readily apparent to those skilled in the art, such as containers containing one or more pharmaceutically acceptable carriers, additional containers, etc. Kits may also include instructions as an insert or as a label indicating the amounts of ingredients to be administered, guidelines for administration, and/or guidelines for mixing the ingredients.

The invention will be described in more detail by specific examples. The following examples are provided for illustrative purposes and are not intended to limit the invention in any way. Those skilled in the art will readily recognize a variety of non-limiting parameters that can be varied or modified to obtain essentially the same results. The compounds of the examples were found to inhibit the activity of KRAS according to at least one assay described herein.

Example

Experimental procedures for compounds of the invention are provided below. Preparative LC-MS purification of a portion of the prepared compounds was performed on Waters mass directed fractionation systems. The basic equipment setup, protocols and control software for operation of these systems have been described in detail in the literature. See, e.g., “Two-Pump At Column Dilution Configuration for Preparative LC-MS”, K. Blom, J. Combi. Chem ., 4, 295 ( 2002 ); “Optimizing Preparative LC-MS Configurations and Methods for Parallel Synthesis Purification”, K. Blom, R. Sparks, J. Doughty, G. Everlof, T. Haque, A. Combs, J. Combi. Chem ., 5, 670 ( 2003 ); and “Preparative LC-MS Purification: Improved Compound Specific Method Optimization”, K. Blom, B. Glass, R. Sparks, A. Combs, J. Combi. Chem ., 6, 874-883 ( 2004 )]. The isolated compounds were typically subjected to analytical liquid chromatography mass spectrometry (LCMS) to check purity.

The isolated compounds were typically subjected to analytical liquid chromatography mass spectrometry (LCMS) for purity check under the following conditions. device; Agilent 1100 Series, LC/MSD, column: Waters Sunfire™ C ₁₈ 5 μm particle size, 2.1 × 5.0 mm, buffer: mobile phase A: 0.025% TFA in water and mobile phase B: acetonitrile; B from 2% to 80% gradient in 3 minutes at a flow rate of 2.0 mL/min.

Some of the prepared compounds were also isolated at production scale by reverse-phase high-performance liquid chromatography (RP-HPLC) or flash chromatography (silica gel) with MS detector as shown in the examples. Typical preparative reversed-phase high-performance liquid chromatography (RP-HPLC) column conditions are:

pH = 2 Purification: Waters Sunfire™ C ₁₈ 5 μm particle size, 19×100 mm column, mobile phase A: 0.1% trifluoroacetic acid (TFA) in water and mobile phase B: eluting with acetonitrile; The flow rate was 30 mL/min and the separation gradient was as described in “Preparative LCMS Purification: Improved Compound Specific Method Optimization”, K. Blom, B. Glass, R. Sparks, A. Combs, J. Comb. Chem. , 6 , 874-883 (2004). Typically, the flow rate used with a 30 x 100 mm column was 60 mL/min.

pH ₌ 10 _Purification : Waters The flow rate was 30 mL/min and the separation gradient was as described in “Preparative LCMS Purification: Improved Compound Specific Method Optimization”, K. Blom, B. Glass, R. Sparks, A. Combs, J. Comb. Chem. , 6 , 874-883 (2004). Typically, the flow rate used with a 30 x 100 mm column was 60 mL/min.

The following abbreviations may be used herein: AcOH (acetic acid); Ac ₂ O (acetic anhydride); aq. (Mercury); atm.(wait(s)); Boc ( t -butoxycarbonyl); BOP ((benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate); br(broad); Cbz (carboxybenzyl); calc.(calculated value); d (double line); dd (double line of double lines); DBU (1,8-diazabicyclo[5.4.0]undec-7-ene); DCM (dichloromethane); DIAD ( N , N' -diisopropyl azidodicarboxylate); DIEA (N,N-diisopropylethylamine); DIPEA ( N , N -diisopropylethylamine); DIBAL (diisobutyl aluminum hydride); DMF ( N , N -dimethylformamide); Et (ethyl); EtOAc (ethyl acetate); FCC (flash column chromatography); g(gram(s)); h(time(s)); HATU ( N , N , N' , N' -tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate); HCl (hydrochloric acid); High-performance liquid chromatography (HPLC); Hz (hertz); J (coupling constant); LCMS (liquid chromatography-mass spectrometry); LDA (lithium diisopropylamide); m(polyline); M(mole); m CPBA (3-chloroperoxybenzoic acid); MS (mass spectrometry); Me (methyl); MeCN (acetonitrile); MeOH (methanol); mg(milligram(s)); min.(minute(s)); mL (millimeter(s)); mmol(millimol(s)); N (normal); NCS (N-chlorosuccinimide); NEt ₃ (triethylamine); nM (nanomole); NMP (N-methylpyrrolidinone); NMR (nuclear magnetic resonance spectroscopy); OTf (trifluoromethanesulfonate); Ph(phenyl); pM (picomole); PPT (precipitate); RP-HPLC (reverse-phase high-performance liquid chromatography); rt (room temperature), s (single line); t (triple or cubic); TBS (tert-butyldimethylsilyl); tert(3rd); tt (triplet of triplets); TFA (trifluoroacetic acid); THF (tetrahydrofuran); μg (microgram(s)); μL (microliter(s)); μM (micromolar); wt% (weight percent). Brine is saturated aqueous sodium chloride. In vacuum ( in vacuo ) is vacuum.

Intermediate 1. 7-Bromo-2,4-dichloro-8-fluoro-6-iodo-3-nitroquinoline

Step 1. 2-Amino-4-bromo-3-fluoro-5-iodobenzoic acid

NIS (9.61 g, 42.7 mmol) was added to a solution of 2-amino-4-bromo-3-fluorobenzoic acid (10.0 g, 42.7 mmol) in DMF (100 mL) and the reaction was then incubated at 80°C for 6 hours. It was stirred for a while. The mixture was cooled with ice water, then water (150 mL) was added and stirred for 20 minutes, and the precipitate was filtered, washed with water, and dried to provide the desired product as a solid. LCMS calculated for C ₇ H ₅ BrFINO ₂ (M+H) ⁺ : m/z = 359.9; Actual value 359.8.

Step 2. 7-Bromo-8-fluoro-6-iodo-2H-benzo[d][1,3]oxazine-2,4(1H)-dione

To a solution of 2-amino-4-bromo-3-fluoro-5-iodobenzoic acid (8.4 g, 23.34 mmol) in 1,4-dioxane (200 mL) was added triphosgene (6.34 g, 21.37 mmol). , and stirred at 100°C for 1 hour. After cooling to room temperature, ice was added until solids precipitated. This mixture was then thoroughly diluted with water (final volume -400 mL) and the solid was collected by filtration and then air dried. The crude product was used in the next step without further purification.

Step 3. 7-Bromo-8-fluoro-6-iodo-3-nitroquinoline-2,4-diol

DIPEA (6.06 ml, 34.7 mmol) was added to a solution of ethyl 2-nitroacetate (4.62 g, 17.36 mmol) in toluene (10.0 mL) at room temperature (rt) and stirred for 10 min. Subsequently, 7-bromo-8-fluoro-6-iodo- 2H -benzo[ d ][1,3]oxazine-2,4( 1H )-dione (6.7g, 17.36 mmol) was added, and the reaction was stirred at 95°C for 3 hours. The reaction was cooled with ice water and then 1N HCl (40 mL) was added. The solid precipitate was collected through filtration and then washed with a small amount of ethyl acetate to provide the desired product as a yellow solid (6 g, 81%). LCMS calculated for C ₉ H ₄ BrFIN ₂ O ₄ (M+H) ⁺ : m/z = 428.8; Actual value 428.8.

Step 4. 7-Bromo-2,4-dichloro-8-fluoro-6-iodo-3-nitroquinoline

DIPEA (3.67 mL, 21.03 mmol) was dissolved in ₇ -bromo-8-fluoro-6-iodo-3-nitroquinoline-2,4-diol (4.51 g, 10.51 mmol) was added to the mixture, and the reaction was then stirred at 105°C for 3 hours. The solvent was removed under vacuum and then azeotroped with toluene three times to give the crude material, which was used in the next step without further purification.

Intermediate 2. tert -Butyl (2 S ,4 S )-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-iodo-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1-yl)-2-(2-( tert -Butoxy)-2-oxoethyl)piperidine-1-carboxylate

Step 1. tert-Butyl (2S,4S)-4-((3-amino-7-bromo-2-(3-(dimethylamino)-3-methylazetidin-1-yl)-8-fluoro-6 -iodoquinolin-4-yl)amino)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate

DIPEA (15.37 ml, 88 mmol) was reacted with 7-bromo-2,4-dichloro-8-fluoro-6-iodo-3-nitroquinoline ( Intermediate 1 ) (20.49) in CH ₂ Cl ₂ (100 mL). g, 44 mmol) and tert -butyl ( 2S , 4S )-4-amino-2-(2-( tert -butoxy)-2-oxoethyl)piperidine-1-carboxylate ( Intermediate 6 ) (13.83g, 44 mmol) was added to the solution at room temperature. The reaction was stirred at 50°C for 3 hours. Upon complete conversion, N,N ,3-trimethylazetidin-3-amine dihydrochloride (9.98 g, 52.8 mmol) and more 2 equivalents of DIPEA (15.37 mL, 88 mmol) were added to the reaction mixture, It was stirred at ℃ overnight. The reaction contents were transferred to a separatory funnel and washed with saturated NH ₄ Cl (200 mL) and brine (100 mL). The organic phase was dried over MgSO ₄ and concentrated.

The concentrated residue was redissolved in MeOH (50 mL), CH ₂ Cl ₂ (10 mL) and aqueous ammonium hydroxide (57 mL, 440 mmol). Sodium dithionite (23 g, 132 mmol) was added in one portion and the reaction was then stirred vigorously at room temperature overnight. Upon completion, the reaction was quenched by addition of H ₂ O (100 mL) and extracted with CH ₂ Cl ₂ (100 mL). The organic phase was washed twice with H ₂ O, dried over Na ₂ SO ₄ and then concentrated to give the desired diamine product as a red viscous oil (18.21 g, 56% over 2 steps). LCMS calculated for C ₃₁ H ₄₆ BrFIN ₆ O ₄ (M+H) ⁺ : m/z = 791.2; Actual value: 791.1.

Step 2. tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-io Do-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1 -Carboxylates

Crude diamine from step 1 (18.21 g, 23 mmol) was dissolved in glacial acetic acid (57.9 mL, 1012 mmol). Sodium nitrite (2.38 g, 34.5 mmol) was added in one portion. The reaction was stirred at room temperature for 2 hours. Upon completion, the contents were poured into vigorously stirred ice water. The precipitated solid was collected through filtration and washed with NaHCO ₃ , water and diethyl ether. The solid was then dried under vacuum to provide the desired product as a brown solid (15 g, 81% yield). LCMS calculated for C ₃₁ H ₄₃ BrFIN ₇ O ₄ (M+H) ⁺ : m/z = 802.2; Actual value 802.1.

Intermediate 3. tert -Butyl (2 S ,4 S )-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate

Step 1. tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-( trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)p Peridine-1-carboxylate

Inject intermediate 2 (1.10 g, 1.37 mmol), copper(I) iodide (39 mg, 0.21 mmol), 1,10-phenanthroline (37 mg, 0.21 mmol), and potassium fluoride (239 mg, 411 mmol) into a pressure vessel. did. DMSO (2.74 mL) was added and the vessel was flushed with N ₂ for 5 minutes. After adding trimethyl borate (0.46 ml, 4.11 mmol) and trimethyl(trifluoromethyl)silane (0.61 mL, 4.11 mmol), the pressure vessel was sealed and heated to 80°C overnight. The container was cooled to room temperature, then cooled in a batch of ice and carefully opened. The reaction mixture was diluted with EtOAc (20 mL), washed with NaHCO ₃ , brine, and concentrated. The crude product was purified on silica gel (20 g, 50-100% EtOAc in CH ₂ Cl ₂ ) to give a brown solid (634 mg, 62% yield). LCMS calculated for C ₃₂ H ₄₃ BrF ₄ N ₇ O ₄ (M+H) ⁺ : m/z = 744.3; Actual value: 744.2.

Step 2. tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-( trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate

tert -Butyl (2 S ,4 S )-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(tri Fluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-2-(2-( tert -butoxy)-2-oxoethyl)p Peridine-1-carboxylate (6.5 g, 8.7 mmol) was dissolved in dioxane (20 mL) and lithium hydroxide solution (7.27 mL, 6M in H ₂ O) was added. The reaction was heated to 80°C overnight. Upon complete hydrolysis of the tert -butyl ester, saturated NH ₄ Cl (100 mL) was added and the reaction was extracted with EtOAc (3 x 100 mL). The combined organic layers were dried over Na ₂ SO ₄ and concentrated to dryness. 2-((2 S ,4 S )-4-(7-bromo-4-(3-(dimethylamino)-3-methylase tidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1- ( tert -butoxycarbonyl)piperidin-2-yl)acetic acid was provided. LCMS calculated for C ₂₈ H ₃₅ BrF ₄ N ₇ O ₄ (M+H) ⁺ : m/z = 688.2; Actual value 688.1.

The carboxylic acid was redissolved in THF (20 mL). DIPEA (3.1 mL, 17.5 mmol) was added and the reaction mixture was cooled to 0°C. Then, isobutyl chloroformate (1.7 mL, 13.1 mmol) was added. After stirring at 0° C. for 20 min, ammonium hydroxide (11.3 mL, 87 mmol) was added and the mixture was stirred for an additional 10 min. Upon completion, the reaction was diluted with EtOAc (20 mL), washed with brine, dried over MgSO ₄ and concentrated to give tert -butyl ( 2S , 4S )-2-(2-amino-2-oxoethyl)- 4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1 H -[1, 2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate was provided. LCMS calculated for C ₂₈ H ₃₆ BrF ₄ N ₈ O ₃ (M+H) ⁺ : m/z = 687.2; Actual value 687.4.

The crude amide was redissolved in THF (20 mL) and cooled to 0°C. Triethylamine (4.9 mL, 34.9 mmol) and TFAA (1.8 mL, 13.1 mol) were added sequentially. After stirring for 1 hour, the reaction was purified with aq. The reaction was quenched by addition of NaHCO ₃ (50 mL), extracted with EtOAc (50 mL), dried over Na ₂ SO ₄ , concentrated and purified on silica (100 g, 0-100% EtOAc in CH ₂ Cl ₂ ) to give the title product. The product was obtained as a yellow solid (4.3 g, 74% yield over 3 steps). LCMS calculated for C ₂₈ H ₃₄ BrF ₄ N ₈ O ₂ (M+H) ⁺ : m/z = 669.2; Actual value 669.4.

Intermediate 4. tert -Butyl (2 S ,4 S )-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1-yl)-2-(2-( tert -Butoxy)-2-oxoethyl)piperidine-1-carboxylate

Step 1. tert-Butyl (2S,4S)-4-(7-bromo-4-chloro-6-fluoro-8-iodo-1H-[1,2,3]triazolo[4,5 -c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate

DIPEA (1.92 ml, 11 mmol) was reacted with 7-bromo-2,4-dichloro-8-fluoro-6-iodo-3-nitroquinoline ( Intermediate 1 ) (2.56) in CH ₂ Cl ₂ (10 mL). g, 5.5 mmol) and tert -butyl ( 2S , 4S )-4-amino-2-(2-( tert -butoxy)-2-oxoethyl)piperidine-1-carboxylate ( Intermediate 6 ) (1.73g, 5.5 mmol) was added to the solution at room temperature. The reaction was stirred at 50°C for 3 hours. Upon complete conversion, the reaction contents were transferred to a separatory funnel and washed with saturated NH ₄ Cl (50 mL) and brine (50 mL). The organic phase was dried over MgSO ₄ and concentrated.

The concentrated residue was redissolved in MeOH (5 mL), CH ₂ Cl ₂ (5 mL) and aqueous ammonium hydroxide (7.3 mL, 55 mmol). Sodium dithionite (2.88 g, 16.5 mmol) was added in one portion and the reaction was then stirred vigorously at room temperature overnight. Upon completion, the reaction was quenched by addition of H ₂ O (10 mL) and extracted with CH ₂ Cl ₂ (50 mL). The organic phase was washed twice with H ₂ O, dried over Na ₂ SO ₄ and then concentrated to provide the desired diamine product.

Crude diamine was dissolved in glacial acetic acid (7.0 mL). Sodium nitrite (0.76 g, 11 mmol) was added in one portion. The reaction was stirred at room temperature for 20 minutes. Upon completion, the reaction contents were poured into vigorously stirred ice water. This reaction mixture was extracted with DCM. The crude product was purified on silica gel (50 g, 0-100% EtOAc in CH ₂ Cl ₂ ) to give the desired product (1.8 g, 70% yield). LCMS calculated for C ₂₅ H ₃₀ BrClFIN ₅ O ₄ (M+H) ⁺ : m/z = 724.0; Actual value: 724.0.

Step 2. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-[1,2,3]triazolo[ 4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate

tert -Butyl (2 S ,4 S )-4-(7-bromo-4-chloro-6-fluoro-8-iodo-1 H -[1,2,3]triazolo[4,5 - c ]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate (1.44 g, 1.99 mmol) was dissolved in CH ₂ Cl ₂ (5 mL). and MeOH (5 mL) and stirred at room temperature until homogeneous. Sodium thiomethoxide (0.28 g, 3.97 mmol) was added in one portion. After stirring for 1 hour, the reaction was quenched with saturated NH ₄ Cl (10 mL), extracted with EtOAc (20 mL), and then washed with NaHCO ₃ . The combined organic layers were dried over MgSO4, filtered and concentrated. LCMS calculated for C ₂₆ H ₃₃ BrFIN ₅ O ₄ S (M+H) ⁺ : m/z = 736.1; Actual value: 736.0.

Intermediate 5. tert -Butyl 4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1-yl)piperidine-1-carboxylate

This compound is obtained by combining tert -butyl ( 2S , 4S )-4-amino-2-(2-( tert -butoxy)-2-oxoethyl)piperidine-1-carboxylate with tert- butyl 4- Prepared according to the procedure described in Intermediate 4 , replacing it with aminopiperidine-1-carboxylate. LCMS calculated for C ₂₀ H ₂₃ BrFIN ₅ O ₂ S (M+H) ⁺ : m/z = 622.0; Actual value: 622.1.

Intermediate 6. tert -Butyl (2 S ,4 S )-4-Amino-2-(2-( tert -Butoxy)-2-oxoethyl)piperidine-1-carboxylate

Step 1. tert-Butyl (R)-6-cyano-5-hydroxy-3-oxohexanoate

A solution of 2.0 M LDA (100 mL, 200 mmol) in anhydrous THF (223 mL) was cooled to -78°C for 1 h, then tert -butyl acetate (26.9 mL, 200 mmol) was added dropwise with stirring over 20 min. Added. After maintaining at -78°C for an additional 40 minutes, a solution of ethyl ( R )-4-cyano-3-hydroxybutanoate (10.5 g, 66.8 mmol) was added dropwise. The mixture was stirred at -40°C for 4 hours, and then an appropriate amount of HCl (2 M) was added to the mixture to maintain pH ~6. During this reaction stop, the temperature of the mixture was maintained at -10°C. Upon completion, the temperature of this mixture was cooled to 0°C. This mixture was extracted with ethyl acetate (3 x 100 mL). The combined organic layers were washed with NaHCO ₃ (100 mL) and brine (100 mL), dried over anhydrous Na ₂ SO ₄ and concentrated to give the material as a yellow oil (15.0 g, 99% yield).

Step 2. tert-Butyl (2S,4R)-2-(2-(tert-butoxy)-2-oxoethyl)-4-hydroxypiperidine-1-carboxylate

A solution of tert-butyl ( R )-6-cyano-5-hydroxy-3-oxohexanoate (15.0 g, 66.0 mmol) in acetic acid (110 mL) was added to platinum(IV) oxide hydrate (0.868 g, 3.30 mmol). ) was treated. The Parr bottle was evacuated, recharged with H ₂ three times, and stirred for 3 hours at 22° C. under H ₂ atmosphere (45 psi, recharged 4 times). The mixture was filtered through Celite and the filter cake was washed with EtOH. The filter cake was concentrated to give the product at a cis:trans diastereomer ratio of ˜9:1.

The residue was dissolved in methanol (100 mL) followed by Boc-anhydride (15.32 ml, 66.0 mmol) and sodium carbonate (13.99 g, 132 mmol) was added. The reaction mixture was stirred overnight at room temperature. This mixture was filtered and concentrated. The residue was purified using a silica gel column to provide the desired product (11.7 g, 56%). LCMS calculated for C ₁₆ H ₂₉ NNaO ₅ (M+Na) ⁺ : m/z = 338.2; Actual value: 338.2.

Step 3. tert-Butyl (2S,4S)-4-azido-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate

tert -butyl (2 S ,4 R )-2-(2-( tert -butoxy)-2-oxoethyl)-4-hydroxypiperidine-1-carboxylate (2.10 g, To a solution of 6.66 mmol), TEA (1.58 ml, 11.32 mmol) and Ms-Cl (0.67 mL, 8.66 mmol) were added at 0°C. After stirring for 1 hour, the reaction was diluted with water and the organic layer was separated, dried over Na ₂ SO ₄ , filtered and concentrated. The obtained residue was dissolved in DMF, sodium azide (1.3 g, 20 mmol) was added, and the reaction mixture was heated at 70° C. for 5 hours. After cooling to room temperature, the reaction was diluted with EtOAc and water. The organic layer was separated, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified using a silica gel column to provide the desired product (1.90 g, 84%). (Product-Boc) LCMS calculated for C ₁₁ H ₂₁ N ₄ O ₂ (M+H) ⁺ : m/z = 241.2; Actual value: 241.2.

Step 4. tert-Butyl (2S,4S)-4-amino-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate

tert -butyl (2 S ,4 S )-4-azido-2-(2-( tert -butoxy)-2-oxoethyl)piperidine-1-carboxylate (1.9 g, To a solution of 5.58 mmol) was added 10% palladium on carbon (0.594 g, 0.558 mmol). The reaction mixture was evacuated under vacuum, recharged with H ₂ and stirred at room temperature for 2 hours. The reaction mixture was filtered through a pad of Celite and washed with methanol. The filtrate was concentrated and used as is (1.5 g, 85%). LCMS calculated for C ₁₆ H ₃₁ N ₂ O ₄ (M+H) ⁺ : m/z = 315.2; Actual value: 315.2.

Intermediate 7. tert -Butyl (2 S ,4 S )-4-Amino-2-(cyanomethyl)piperidine-1-carboxylate

Step 1. tert-Butyl (2S,4S)-4-(((benzyloxy)carbonyl)amino)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxyl rate

To a stirred solution of intermediate 6 (5.2 g, 16.54 mmol) in CH ₂ Cl ₂ (80.0 mL) was added N -(benzyloxy-carbonyloxy)succinimide (4.95 g, 19.85 mmol) followed by DIPEA (4.33 mL, 24.81 mmol) was added. The mixture was stirred at room temperature for 2 hours and then diluted with water. This mixture was extracted with water and brine. The combined organic layers were dried over MgSO ₄ , filtered, concentrated and used directly in the next step without further purification.

Step 2. 2-((2S,4S)-4-(((benzyloxy)carbonyl)amino)piperidin-2-yl)acetic acid

The concentrated residue from Step 1 was charged in CH ₂ Cl ₂ (80.0 mL) and TFA (50.0 mL). This mixture was stirred at room temperature overnight, then concentrated to dryness and used directly in the next step without further purification.

Step 3. 2-((2S,4S)-4-(((benzyloxy)carbonyl)amino)-1-(tert-butoxycarbonyl)piperidin-2-yl)acetic acid

The concentrated residue from step 2 was charged in CH ₂ Cl ₂ (80.0 mL) and triethylamine (23.1 mL, 165 mmol) was added slowly. The mixture was stirred at room temperature for 5 minutes, then Boc-anhydride (4.33 g, 19.85 mmol) was added. The mixture was stirred at room temperature for a further 30 minutes. Additional Boc-anhydride was added as needed. Upon complete conversion, the mixture was acidified to pH 4-5 and then extracted with EtOAc. The combined organic layers were dried over MgSO ₄ , filtered, concentrated and used directly in the next step without further purification.

Step 4. tert-Butyl (2S,4S)-2-(2-amino-2-oxoethyl)-4-(((benzyloxy)carbonyl)amino)piperidine-1-carboxylate

The concentrated residue from step 3 was charged in THF (80.0 mL) and DIPEA (8.67 mL, 49.6 mmol). The mixture was cooled to 0°C, and then isobutyl chloroformate (5.43 mL, 41.3 mmol) was added slowly. The mixture was stirred at 0° C. for a further 20 minutes, then ammonium hydroxide (28% in water, 23.0 mL, 165 mmol) was added to the mixture. After stirring for a further 5 minutes, the mixture was diluted with brine and extracted with EtOAc. The combined organic layers were dried over MgSO ₄ , filtered, concentrated and purified by column chromatography (0-8% MeOH in DCM).

Step 5. tert-Butyl (2S,4S)-4-(((benzyloxy)carbonyl)amino)-2-(cyanomethyl)piperidine-1-carboxylate

The purified product from step 4 was charged in THF (80.0 mL) and triethylamine (6.0 mL, 43 mmol). The mixture was cooled to 0°C, and then TFAA (3.5 mL, 24.8 mmol) was added slowly. The mixture was stirred at 0° C. for a further 30 minutes, then diluted with EtOAc and extracted with brine. The combined organic layers were dried over MgSO ₄ , filtered, concentrated to dryness and purified on silica gel to give the desired product (5.12 g, 83% over 5 steps). LCMS calculated for C ₁₆ H ₂₀ N ₃ O ₄ (M+H-tert-butyl) ⁺ : m/z = 318.1; Actual value: 318.1.

Step 6. tert-Butyl (2S,4S)-4-amino-2-(cyanomethyl)piperidine-1-carboxylate

In a round-bottomed flask containing a stir bar, tert -butyl (2 S ,4 S )-4-(((benzyloxy)carbonyl)amino)-2-(cyanomethyl)piperidine-1-carboxylate ( 5.12 g, 13.71 mmol), palladium on carbon (10 wt%, 2.92 g, 2.74 mmol) and MeOH (45 mL) were injected. The round bottom flask was evacuated and backfilled with H ₂ (this process was repeated a total of 3 times), and the mixture was stirred vigorously at room temperature for 1.5 hours with the H ₂ balloon attached. The mixture was then filtered over Celite and the solid was washed with EtOAc. The filtrate was concentrated and used directly in the next step without further purification.

Intermediate 8. tert -Butyl (2 S ,4 S )-4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate

Step 1. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-[1,2,3]triazolo[4,5 -c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate

DIPEA (5.62 ml, 32 mmol) was dissolved in 7-bromo-2,4-dichloro-8-fluoro-6-iodo-3-nitroquinoline ( Intermediate 1 ) (10 g, 21.5 mmol) in MeCN (100 mL). ) and tert -butyl (2 S , 4 S ) -4-amino-2- (cyanomethyl) piperidine-1-carboxylate ( Intermediate 7) (5.15 g, 21.5 mmol) was added at room temperature. . The reaction was stirred at 50°C for 3 hours. Upon complete conversion, the reaction mixture was concentrated.

The residue was dissolved in MeOH (80 mL), sodium thiomethoxide (3.0 g, 43 mmol) was added in one portion at room temperature and upon complete conversion, the reaction mixture was diluted with water, filtered and washed with water to give a brown solid. provided.

The solid was redissolved in MeOH (80 mL), CH ₂ Cl ₂ (20 mL) and aqueous ammonium hydroxide (28 mL, 440 mmol). Sodium dithionite (11.2 g, 132 mmol) was added in one portion and the reaction was then stirred vigorously at room temperature. Upon completion, the reaction was quenched by addition of H ₂ O (100 mL) and extracted with CH ₂ Cl ₂ (100 mL). The organic phase was washed twice with H ₂ O, dried over Na ₂ SO ₄ and then concentrated to give the desired diamine product as a red viscous oil.

Crude diamine was dissolved in glacial acetic acid (50 mL, 875 mmol). Sodium nitrite (2.97 g, 43.0 mmol) was added in one portion. The reaction was stirred at room temperature for 2 hours. Upon completion, the reaction contents were poured into vigorously stirred ice water. The precipitated solid was collected through filtration and washed with NaHCO ₃ , water and diethyl ether. The solid was then dried under vacuum to provide the desired product as a brown solid (7.1 g, 50% yield). LCMS calculated for C ₂₂ H ₂₄ BrFIN ₆ O ₂ S (M+H) ⁺ : m/z = 661.0; Actual value: 660.9.

Step 2. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4 ,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate

tert -butyl (2 S ,4 S )-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1 H- in a screw-cap vial equipped with a magnetic stir bar. [1,2,3]triazolo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate (1.19g, 1.832 mmol), methylboronic acid (1.096 g, 18.32 mmol), tripotassium phosphate (1.166 g, 5.49 mmol) and bis(triphenylphosphine)-palladium(II) chloride (257 mg, 0.366 mmol) followed by dioxane (10.0 mL) and water (2.0 mL). ) was injected. The vial was sealed with a Teflon-lined septum, evacuated, and refilled with nitrogen (this process was repeated three times). The reaction was then stirred at 90°C for 3 hours. After cooling to room temperature, the mixture was diluted with brine and extracted with EtOAc. The combined organic layers were dried over MgSO ₄ , filtered, concentrated to dryness and purified on silica gel to give the desired product. LCMS calculated for C ₂₃ H ₂₇ BrFN ₆ O ₂ S (M+H) ⁺ : m/z = 549.1; Actual value: 549.1.

Example 1a and Example 1b. 2-((2 S ,4 S )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(tri fluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1-yl)-1-(( E )-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile

Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro- 7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperi Din-1-carboxylate

Intermediate 3 (1.3g, 1.9 mmol), XPhos Pd G2 (76mg, 0.097 mmol), (5-fluoroquinolin-8-yl)boronic acid (408mg, 2.1 mmol), K ₃ PO ₄ (1.24g) in a reaction vial. , 5.83 mmol) and dioxane (5 mL) and H ₂ O (1 mL) were injected. This mixture was sparged with N ₂ for 5 minutes, then heated to 90° C. and stirred at this temperature for 1 hour. Upon completion, the reaction was cooled to room temperature, diluted with EtOAc (20 mL), and aq. Washed with NH ₄ Cl (20 mL). The organic phase was separated, dried over MgSO ₄ , filtered and then concentrated. The crude product was first purified by silica (20 g, 50-100% EtOAc in CH ₂ Cl ₂ ) and then purified by preparative-LCMS (XBridge C18 column, flow rate of 60 mL/min, containing 0.15% NH ₄ OH). Further purification using an acetonitrile/water gradient) provided the desired product as a pair of diastereomers (white amorphous powder, 21% combined yield).

Diastereomer 1. Peak 1. LCMS calculated for C ₃₇ H ₃₉ F ₅ N ₉ O ₂ (M+H) ⁺ m/z = 736.3; Actual value 736.2.

Diastereomer 2. Peak 2. LCMS calculated for C ₃₇ H ₃₉ F ₅ N ₉ O ₂ (M+H) ⁺ m/z = 736.3; Actual value 736.2.

Step 2. 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinoline -8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4- Fluorobut-2-enoyl)piperidin-2-yl)acetonitrile

tert -butyl (2 S ,4 S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6- from Step 1 Fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1- 1) TFA (1 mL) was added to the reaction vial injected with piperidine-1-carboxylate (diastereomer 1, 150 mg, 0.24 mmol) at room temperature. After stirring for 15 minutes, volatile substances were removed. The residue was redissolved in acetonitrile (2 mL) and cooled to 0°C. To this reaction was added DIPEA (0.36 mL), followed by ( E )-4-fluorobut-2-enoic acid (42 mg, 0.41 mmol) and anhydrous propylphosphonic acid solution (50% in EtOAc, 0.25 mL, 0.41 mmol). ) was added. After stirring at 0°C for 10 min, the reaction was incubated with aq. The reaction was quenched with NaHCO ₃ (5 mL), extracted with EtOAc (5 mL), washed with brine and concentrated. The crude product was redissolved in acetonitrile and purified using prep-LCMS (XBridge C18 column, eluting with an acetonitrile/water gradient containing 0.1% TFA at a flow rate of 60 mL/min) to give Example 1a. (Diastereomer 1) was provided as the TFA salt in the form of a white amorphous powder (50 mg free equivalent, 34% yield). LCMS calculated for C ₃₆ H ₃₄ F ₆ N ₉ O (M+H) ⁺ m/z = 722.3; Actual value 722.2. ^1H NMR (500 MHz, DMSO- d6 ) δ 10.80 (s, 1H), 8.87 (dd, J = 4.2, 1.7 Hz, 1H), 8.62 (dd, J = 8.5, 1.7 Hz, 1H), 8.41 ( s, 1H), 7.82 (dd, J = 8.0, 5.8 Hz, 1H), 7.70 (dd, J = 8.5, 4.2 Hz, 1H), 7.65 (dd, J = 9.8, 8.0 Hz, 1H), 6.90 - 6.77 (m, 2H), 5.93 (s, 1H), 5.32 (s, 1H), 5.24 - 5.20 (m, 1H), 5.12 (dd, J = 3.6, 1.2 Hz, 1H), 4.97 (s, 1H), 4.76 (s, 1H), 4.57 (s, 1H), 4.37 - 4.21 (m, 2H), 3.61 (d, J = 12.7 Hz, 1H), 3.39 - 3.21 (m, 2H), 2.84 (s, 6H) , 2.46 - 2.22 (m, 4H), 1.70 (s, 3H).

Example 1b (diastereomer 2) is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3- from Step 1 Methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo [4,5- c ]quinolin-1-yl)piperidine-1-carboxylate (diastereomer 1) from Step 1 with tert -butyl (2 S ,4 S )-2-(cyanomethyl )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(tri Replace with fluoromethyl) -1H- [1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate (diastereomer 2) in the above procedure. It was manufactured using . LCMS calculated for C ₃₆ H ₃₄ F ₆ N ₉ O (M+H) ⁺ m/z = 722.3; Actual value 722.2.

Example 2a and Example 2b. 2-((2 S ,4 S )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(2-methoxy-3-methylphenyl)-8-(trifluoro Romethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1-yl)-1-(( E )-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile

Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-( 2-methoxy-3-methylphenyl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1- carboxylate

Intermediate 3 (100 mg, 0.15 mmol), (2-methoxy-3-methylphenyl)boronic acid (30 mg, 0.18 mmol), Pd(PPh ₃ ) ₄ (17 mg, 0.015 mmol), K ₃ PO ₄ (95 mg) in a reaction vial. , 0.45 mmol) and dioxane (2.0 mL) and H ₂ O (0.4 mL) were injected. This mixture was sparged with N ₂ for 5 minutes and then heated at 90° C. for 1 hour. Upon completion, the reaction was cooled to room temperature, diluted with EtOAc (5.0 mL), and aq. Washed with NH ₄ Cl (5.0 mL). The organic phase was separated, dried over MgSO ₄ and concentrated. The crude product was purified by silica (10 g, 50-100% EtOAc in DCM) to provide the desired product as a pale yellow viscous oil (92 mg, 87% yield). LCMS calculation for C ₃₆ H ₄₃ F ₄ N ₈ O ₃ (M+H) ⁺ m/z = 711.3; Actual value 711.2.

Step 2. 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(2-methoxy- 3-methylphenyl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluo Robut-2-enoyl)piperidin-2-yl)acetonitrile

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6- Fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1- 1) piperidine-1-carboxylate tert -butyl (2 S ,4 S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidine- 1-yl)-6-fluoro-7-(2-methoxy-3-methylphenyl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate, prepared according to the procedure described in Example 1a and Example 1b , Step 2 .

Example 2a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₅ H ₃₈ F ₅ N ₈ O ₂ (M+H) ⁺ : m/z = 697.3; Actual value: 697.3.

Example 2b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₅ H ₃₈ F ₅ N ₈ O ₂ (M+H) ⁺ : m/z = 697.3; Actual value: 697.3.

Example 3a and Example 3b. 2-((2 S ,4 S )-4-(7-(3-chloro-2-methoxyphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(tri fluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1-yl)-1-(( E )-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile

Step 1. tert-Butyl (2S,4S)-4-(7-(3-chloro-2-methoxyphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6- Fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1 -Carboxylates

This compound was prepared following the procedure described in Examples 2a and 2b , Step 1 , by replacing (2-methoxy-3-methylphenyl)boronic acid with (3-chloro-2-methoxyphenyl)boronic acid. did. LCMS calculated for C ₃₅ H ₄₀ ClF ₄ N ₈ O ₂ (M+H) ⁺ : m/z = 731.3; Actual value: 731.2.

Step 2. 2-((2S,4S)-4-(7-(3-chloro-2-methoxyphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl) -6-fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4- Fluorobut-2-enoyl)piperidin-2-yl)acetonitrile

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6- Fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1- 1) piperidine-1-carboxylate tert -butyl (2 S ,4 S )-4-(7-(3-chloro-2-methoxyphenyl)-4-(3-(dimethylamino)- 3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5 -c ]quinolin-1-yl )-2-(cyanomethyl)piperidine-1-carboxylate, prepared according to the procedure described in Example 1a and Example 1b , Step 2 .

Example 3a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₄ H ₃₅ ClF ₅ N ₈ O ₂ (M+H) ⁺ : m/z = 717.3; Actual value: 717.2.

Example 3b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₄ H ₃₅ ClF ₅ N ₈ O ₂ (M+H) ⁺ : m/z = 717.3; Actual value: 717.2.

Example 4a and Example 4b. 2-((2 S ,4 S )-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-(( S )-One-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1-yl)-1-(( E )-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile

Step 1. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-4-(methylthio)-8-(trifluoromethyl)-1H-[1,2,3]triazolo [4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate

Intermediate 4 (3.50 g, 4.75 mmol) and methyl 2,2-difluoro-2-(fluorosulfonyl)-acetate (1.21 ml, 9.51 mmol) (MFDA), copper(I) iodide (0.272 mmol) in a sealed tube. g, 1.426 mmol) and NMP (20 mL) were injected. The tube was flushed with N ₂ and then sealed and heated at 80° C. overnight. The reaction was then cooled to room temperature, poured into NaHCO ₃ and extracted with EtOAc. The organic layers were combined, dried over MgSO ₄ and concentrated. The crude product was purified on silica (40 g, 0-50% EtOAc in hexane) to give the desired product (2.5 g, 78% yield). LCMS calculated for C ₂₇ H ₃₃ BrF ₄ N ₅ O ₄ S (M+H) ⁺ : m/z = 678.1; Actual value: 678.3.

Step 2. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-4-(methylthio)-8-(trifluoromethyl)-1H-[1,2,3]triazolo [4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate

This compound is tert -butyl ( 2S , 4S )-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro- 8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-2-(2-( tert -butoxy)-2- Oxoethyl)piperidine-1-carboxylate was reacted with tert -butyl ( 2S , 4S )-4-(7-bromo-6-fluoro-4-(methylthio)-8-(trifluoromethyl )-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-2-(2-( tert -butoxy)-2-oxoethyl)piperidine- Replaced with 1-carboxylate, it was prepared according to the procedure described in Intermediate 3 , Step 2 . LCMS calculated for C ₂₃ H ₂₄ BrF ₄ N ₆ O ₂ S (M+H) ⁺ : m/z = 603.1; Actual value: 603.2.

Step 3. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-(methylthio)-8-(tri Fluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate

This compound replaces intermediate 3 with tert -butyl (2 S ,4 S )-4-(7-bromo-6-fluoro-4-(methylthio)-8-(trifluoromethyl)-1 H - [1,2,3]triazolo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate, Example 1a and Example 1b , prepared according to the procedure described in Step 1 . LCMS calculated for C ₃₂ H ₂₉ F ₅ N ₇ O ₂ S (M+H) ⁺ : m/z = 670.2; Actual value: 670.2.

Step 4. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-(( S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinoline-1- 1) Piperidine-1-carboxylate

tert -Butyl (2 S ,4 S )-2-(cyanomethyl)-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-(methylthio)-8- (Trifluoromethyl) -1H- [1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate (85 mg, 0.13 mmol) was reacted with CH ₂ Dissolve in Cl ₂ (1 mL) and cool to 0°C. mCPBA (33 mg, 0.19 mmol) was added in one portion and the reaction was stirred for 30 minutes and then quenched by the addition of saturated NaHCO ₃ (2 mL). This mixture was extracted with CH ₂ Cl ₂ (5 mL). The combined organic layers were dried over MgSO ₄ , filtered, and then concentrated to provide a mixture of crude sulfoxides and sulfones. The vial filled with ( S )-1-(( S )-1-methylpyrrolidin-2-yl)ethan-1-ol (16.4 mg, 0.13 mmol) and dry THF (1 mL) was cooled to 0°C. . LiHMDS (0.13 mL, 1M in THF) was added. After stirring for 10 minutes, the reaction was added dropwise to a THF solution of crude sulfoxide. After stirring for another 10 minutes, the reaction was quenched by adding saturated NH ₄ Cl (5 mL) and extracted with EtOAc (15 mL). The combined organic layers were dried over MgSO ₄ and purified on silica (0-100% EtOAc in hexanes) to provide the desired product as a white solid (75 mg, 79% yield). LCMS calculated for C ₃₈ H ₄₀ F ₅ N ₈ O ₃ (M+H) ⁺ : m/z = 751.3; Actual value: 751.5.

Step 5. 2-((2S,4S)-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrroli din-2-yl) ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E )-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6- Fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1- 1) Piperidine-1-carboxylate is tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)- 4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo Replaced with [4,5-c]quinolin-1-yl)piperidine-1-carboxylate , prepared according to the procedure described in Example 1a and Example 1b , Step 2 .

Example 4a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₇ H ₃₅ F ₆ N ₈ O ₂ (M+H) ⁺ : m/z = 737.3; Actual value: 737.2.

Example 4b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₇ H ₃₅ F ₆ N ₈ O ₂ (M+H) ⁺ : m/z = 737.3; Actual value: 737.2.

Example 5a and Example 5b. 1-(4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-(( S )-One-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidin-1-yl)prop-2-en-1-one

Step 1. tert-Butyl 4-(7-bromo-6-fluoro-4-(methylthio)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c ]quinoline-1-yl)piperidine-1-carboxylate

This compound was prepared following the procedure described in Examples 4a and 4b , Step 1 , replacing Intermediate 4 with Intermediate 5 . LCMS calculated for C ₂₁ H ₂₃ BrF ₄ N ₅ O ₂ S (M+H) ⁺ : m/z = 564.1; Actual value: 564.0.

Step 2. tert-Butyl 4-(7-bromo-6-fluoro-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoro Romethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-(methylthio )-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate with tert -butyl 4 -(7-Bromo-6-fluoro-4-(methylthio)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline- 1-yl)piperidine-1-carboxylate, prepared according to the procedure described in Examples 4a and 4b , step 4 . LCMS calculated for C ₂₇ H ₃₄ BrF ₄ N ₆ O ₃ (M+H) ⁺ : m/z = 645.2; Actual value: 645.4.

Step 3. tert-Butyl 4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl) Ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate

This compound replaces intermediate 3 with tert -butyl 4-(7-bromo-6-fluoro-4-(( S )-1-(( S )-1-methylpyrrolidin-2-yl) ethoxy )-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate . Prepared according to the procedure described in Example 1a and Example 1b , Step 1 . LCMS calculated for C ₃₆ H ₃₉ F ₅ N ₇ O ₃ (M+H) ⁺ : m/z = 712.3; Actual value: 712.5.

Step 4. 1-(4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2 -yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-1-yl)prop p-2-n-1-on

tert -Butyl 4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-(( S )-1-(( S )-1-methylpyrrolidin-2-yl) Ethoxy)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate (25 mg, TFA (0.5 mL) was added to 0.039 mmol) and stirred at room temperature for 10 minutes. The solvent was removed under vacuum. The residue was dissolved in CH ₂ Cl ₂ (1.0 mL), cooled to 0° C., and to this was added triethylamine (16 μL, 0.116 mmol) followed by acryloyl chloride (5.3 mg, 0.058 mmol), The reaction was stirred at 0°C for 20 minutes. The reaction was diluted with CH ₂ Cl ₂ , washed with saturated NaHCO ₃ , the organic solvent was dried and concentrated. The crude product was redissolved in acetonitrile and purified using preparative-LCMS (XBridge C18 column, eluting with an acetonitrile/water gradient containing 0.1% TFA at a flow rate of 60 mL/min).

Example 5a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₄ H ₃₃ F ₅ N ₇ O ₂ (M+H) ⁺ : m/z = 666.3; Actual value: 666.4.

Example 5b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₄ H ₃₃ F ₅ N ₇ O ₂ (M+H) ⁺ : m/z = 666.3; Actual value: 666.4.

Example 6. 2-((2 S ,4 S )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1-yl)-1-(( E )-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile

Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methylthio)- 1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate

Intermediate 8 (400 mg, 0.728 mmol), (2,3-dimethylphenyl)boronic acid (164 mg, 1.092 mmol), potassium phosphate (464 mg, 2.184 mmol) and Pd(Ph _{3 P) in dioxane (6 mL)/water (1} mL) ) The mixture of ₄ (84 mg, 0.073 mmol) was evacuated and recharged with nitrogen (this process was repeated three times). The reaction was stirred at 105°C for 1 hour. This mixture was diluted with ethyl acetate and washed with water and brine. The organics were filtered, dried and concentrated. The product was purified by column eluting with hexane/EtOAc (maximum EtOAc 80%). LCMS calculated for C ₃₁ H ₃₆ FN ₆ O ₂ S (M+H) ⁺ m/z = 575.3; Actual value 575.3.

Step 2. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methylsulfinyl) -1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate

m-CPBA (167 mg, 0.966 mmol) was reacted with tert -butyl (2 S ,4 S )-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl) in CH ₂ Cl ₂ (5.0 mL). )-6-fluoro-8-methyl-4-(methylthio)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1- A solution of the carboxylate (370.0 mg, 0.644 mmol) was added at 0° C. and the reaction was then stirred at this temperature for 10 minutes. The reaction was quenched by adding saturated Na ₂ S ₂ O ₃ , diluted with ethyl acetate, washed with saturated NaHCO ₃ , brine, filtered, dried, concentrated and the crude was used directly in the next step. . LCMS calculated for C ₃₁ H ₃₆ FN ₆ O ₃ S (M+H) ⁺ m/z = 591.3; Actual value 591.3.

Step 3. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2, 3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate

Triethylamine (0.625 mL, 4.48 mmol) was added to tert -butyl (2 S ,4 S )-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8 -Methyl-4-(methylsulfinyl) -1H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate (0.680g, 1.121 mmol) and N , N ,3-trimethylazetidin-3-amine (0.192 g, 1.681 mmol), and then stirred at 70°C for 2 hours. The product was purified by column eluting with CH ₂ Cl ₂ /MeOH (maximum MeOH 15%). The atropisomers were separated by SFC (column, Phenomenex Lux 5um Cellulose-1, 21.2x250mm, mobile phase 25% MeOH in CO ₂ at 70 mL/min isoc) to give two peaks named PK1 and PK2. LCMS calculated for C ₃₆ H ₄₆ FN ₈ O ₂ (M+H) ⁺ m/z = 641.4; Actual value 641.5.

Step 4. 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6- Fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl ) piperidin-2-yl) acetonitrile

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6- Fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1- 1) Piperidine-1-carboxylate is tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidine-1- 1)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperi replacing dine-1-carboxylate ( PK1 from step 3) and replacing ( E )-4-fluorobut-2-enoic acid with (E)-4-methoxybut-2-enoic acid . Prepared according to the procedure described in Example 1a and Example 1b , Step 2 . LCMS calculated for C ₃₆ H ₄₄ FN ₈ O ₂ (M+H) ⁺ m/z = 639.4; Actual value 639.5. ¹ H NMR (600 MHz, DMSO) δ 8.04 (s, 1H), 7.30 - 7.21 (m, 2H), 7.00 (dd, J = 7.4, 1.5 Hz, 1H), 6.81 - 6.70 (m, 2H), 5.30 (s, 1H), 4.96 (s, 1H), 4.74 (d, J = 13.8 Hz, 1H), 4.28 (d, J = 14.3 Hz, 1H), 4.11 (s, 2H), 3.64 (s, 1H) , 3.39 (dd, J = 17.0, 8.5 Hz, 1H), 3.33 (s, 3H), 3.31 - 3.19 (m, 3H), 2.83 (s, 6H), 2.43 (s, 3H), 2.41 (m, 1H) ) 2.35 (s, 3H), 2.20 (s, 3H), 2.13 (d, J = 13.2 Hz, 1H), 1.94 (s, 3H), 1.70 (s, 3H).

Example 7. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-((S)-1 -((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-( (E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile

Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4 -(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate

This compound is made by combining (2,3-dimethylphenyl)boronic acid with 1-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H- Replaced with indazole, prepared according to the procedure described in Example 6, Step 1 . LCMS calculated for C ₃₁ H ₃₄ FN ₈ O ₂ S (M+H) ⁺ m/z = 601.3; Actual value 601.3.

Step 2. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4 -(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methyl thio)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate with tert -butyl (2 S ,4 S )-2 -(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1 H -indazol-6-yl)-4-(methylthio)-1 H -[1, 2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate was replaced with the procedure described in Example 6, Step 2 . LCMS calculated for C ₃₁ H ₃₄ FN ₈ O ₃ S (M+H) ⁺ m/z = 617.2; Actual value 617.4.

Step 3. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4 -((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinoline-1 -1) Piperidine-1-carboxylate

LiHMDS (1.0 M in THF) (40.7 mg, 0.243 mmol) was dissolved in ( S )-1-(( S )-1-methylpyrrolidin-2-yl)ethan-1-ol (31.4) in THF (1.0 mL). mg, 0.243 mmol) was added to the solution at 0°C and stirred for 5 minutes. The resulting solution was purified in THF (1.0 mL) with tert -butyl (2 S ,4 S )-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1 H- indazol-6-yl)-4-(methylsulfinyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate (75.0 mg, 0.122 mmol) was added at 0°C, and the reaction was then stirred at room temperature for 1 hour. This mixture was diluted with ethyl acetate, washed with saturated NaHCO ₃ , water, filtered, concentrated and used directly in the next step. LCMS calculated for C ₃₇ H ₄₅ FN ₉ O ₃ (M+H) ⁺ m/z = 682.4; Actual value 682.5.

Step 4. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-((S)-1- ((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(( E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6- Fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1- 1) piperidine-1-carboxylate tert -butyl (2 S ,4 S )-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1 H -indazol-6-yl)-4-(( S )-1-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]tri Replaced with azolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate, prepared according to the procedure described in Example 1a and Example 1b , Step 2 . LCMS calculation for C ₃₆ H ₄₀ F ₂ N ₉ O ₂ (M+H) ⁺ m/z = 668.3; Actual value 668.5. ^1H NMR (600 MHz, DMSO) δ 8.22 (s, 1H), 8.17 (s, 1H), 7.93 (d, J = 8.3 Hz, 1H), 7.72 (s, 1H), 7.13 (dd, J = 8.2 Hz, 1H), 6.92 - 6.79 (m, 2H), 5.87 (d, J = 16.1 Hz, 1H), 5.58 (d, J = 7.5 Hz, 1H), 5.31 (d, J = 6.4 Hz, 1H), 5.21 (d, J = 2.4 Hz, 1H), 5.14 (d, J = 2.9 Hz, 2H), 5.00 (s, 1H), 4.76 (d, J = 13.8 Hz, 1H), 4.32 (d, J = 14.2 Hz, 1H), 4.11 (s, 3H), 3.96 (p, J = 7.7 Hz, 1H), 3.65 - 3.59 (m, 3H), 3.45 - 3.23 (m, 2H), 3.19 (m,1H), 3.14 (s, 3H), 2.44 (s, 3H), 2.32- 2.16 (m, 2H), 2.11- 1.89 (m, 2H), 1.59 (s, 3H).

Example 8. 2-((2 S ,4 S )-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-(( S )-One-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1-yl)-1-(( E )-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile

Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-(methylthio )-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate

This compound was prepared following the procedure described in Example 6, Step 1 , replacing (2,3-dimethylphenyl)boronic acid with (6-methylpyridin-3-yl)boronic acid. LCMS calculated for C ₂₉ H ₃₃ FN ₇ O ₂ S (M+H) ⁺ m/z = 562.2; Actual value 562.3.

Step 2. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-(methylsulfy Nyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methyl thio)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate with tert -butyl (2 S ,4 S )-2 -(cyanomethyl)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-(methylthio)-1 H -[1,2,3]tri Azolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate was prepared following the procedure described in Example 6, Step 2 . LCMS calculated for C ₂₉ H ₃₃ FN ₇ O ₃ S (M+H) ⁺ m/z = 578.2; Actual value 578.4.

Step 3. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-((S )-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)p Peridine-1-carboxylate

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl- 1H- indazole-6- 1)-4-(methylsulfinyl) -1H- [1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate was reacted with tert -butyl ( 2 S ,4 S )-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-(methylsulfinyl)-1 H -[1,2,3]Triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate was prepared following the procedure described in Example 7, Step 3 . LCMS calculated for C ₃₅ H ₄₄ FN ₈ O ₃ (M+H) ⁺ m/z = 643.4; Actual value 643.5.

Step 4. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-((S)-1-((S) -1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4 -Fluorobut-2-enoyl)piperidin-2-yl)acetonitrile

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6- Fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1- 1) piperidine-1-carboxylate tert -butyl (2 S ,4 S )-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(6-methylpyridine- 3-yl)-4-(( S )-1-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4, 5- c ]quinolin-1-yl)piperidine-1-carboxylate, prepared according to the procedure described in Example 1a and Example 1b , Step 2 . LCMS calculation for C ₃₄ H ₃₉ F ₂ N ₈ O ₂ (M+H) ⁺ m/z = 629.3; Actual value 629.5.

Example 9. 2-((2 S ,4 S )-4-(6-fluoro-8-methyl-7-(1-methyl-1 H -indazole-3-yl)-4-(( S )-One-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1-yl)-1-(( E )-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile

Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-3-yl)-4 -(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate

This compound is made by combining (2,3-dimethylphenyl)boronic acid with 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H- Replaced with indazole, prepared according to the procedure described in Example 6, Step 1 . LCMS calculated for C ₃₁ H ₃₄ FN ₈ O ₂ S (M+H) ⁺ m/z = 601.3; Actual value 601.1.

Step 2. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-3-yl)-4 -(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methyl thio)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate with tert -butyl (2 S ,4 S )-2 -(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1 H -indazol-3-yl)-4-(methylthio)-1 H -[1, 2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate was replaced with the procedure described in Example 6, Step 2 . LCMS calculated for C ₃₁ H ₃₄ FN ₈ O ₃ S (M+H) ⁺ m/z = 617.3; Actual value 617.3.

Step 3. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-3-yl)-4 -((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinoline-1 -1) Piperidine-1-carboxylate

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl- 1H- indazole-6- 1)-4-(methylsulfinyl) -1H- [1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate was reacted with tert -butyl ( 2 S ,4 S )-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1 H -indazol-3-yl)-4-(methylsulfy Nyl) -1H- [1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate, and follow the procedure described in Step 3 of Example 7. It was prepared accordingly. LCMS calculated for C ₃₇ H ₄₅ FN ₉ O ₃ (M+H) ⁺ m/z = 682.4; Actual value 682.4

Step 4. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-3-yl)-4-((S)-1- ((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(( E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7- (2,3-dimethylphenyl)-6-fluoro-8-methyl-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1- The carboxylate is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl- 1H- indazol-3-yl )-4-(( S )-1-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate, prepared according to the procedure described in Example 6 , Step 4 . LCMS calculated for C ₃₇ H ₄₃ FN ₉ O ₃ (M+H) ⁺ m/z = 680.4; Actual value 680.3.

Example 10. 2-((2 S ,4 S )-4-(6-fluoro-7-(4-fluorophenyl)-8-methyl-4-(( S )-One-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1-yl)-1-(( E )-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile

This compound is made by combining 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole with (4-fluorophenyl)boron. Replaced with acid and prepared according to the procedure described in Example 9 , steps 1 to 4 . LCMS calculated for C ₃₅ H ₄₀ F ₂ N ₇ O ₃ (M+H) ⁺ m/z = 644.3; Actual value 644.4. ¹ H NMR (500 MHz, DMSO) δ 10.09 - 9.21 (s, 1H), 8.51 - 7.69 (s, 1H), 7.53 - 7.44 (dd, J = 8.5, 5.7 Hz, 2H), 7.44 - 7.36 (t, J = 8.9 Hz, 2H), 6.83 - 6.63 (m, 2H), 5.92 - 5.72 (m, 1H), 5.66 - 5.47 (m, 1H), 5.41-4.20 (m, 2H), 4.18 - 4.03 (d, J = 2.5 Hz, 2H), 4.00 - 3.85 (m, 1H), 3.69 - 3.53 (m, 2H), 3.39 - 3.32 (s, 3H), 3.27 - 3.10 (m, 5H), 2.55 - 2.36 (d, J = 13.8 Hz, 6H), 2.38 - 2.23 (m, 2H), 2.19 - 2.04 (m, 2H), 2.00 - 1.88 (m, 2H), 1.64 - 1.50 (d, J = 6.1 Hz, 3H).

Example 11a and Example 11b. 8-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-(( S )-One-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]Quinoline-7-yl)-1-naphtonitrile

Step 1. tert-Butyl 4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinoline -1-yl)piperidine-1-carboxylate

This compound is tert -butyl (2 S ,4 S )-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1 H -[1,2,3] Triazolo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate was replaced with Intermediate 5 and prepared according to the procedure described in Step 2 of Intermediate 8. . LCMS calculated for C ₂₁ H ₂₆ BrFN ₅ O ₂ S (M+H) ⁺ m/z = 510.1; Actual value 510.1.

Step 2. tert-Butyl 4-(7-bromo-6-fluoro-8-methyl-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c] Quinoline-1-yl)piperidine-1-carboxylate

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methyl thio)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate with tert -butyl 4-(7-bromo-6 -fluoro-8-methyl-4-(methylthio)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate Alternatively, it was prepared according to the procedure described in Example 6, Step 2 . LCMS calculated for C ₂₁ H ₂₆ BrFN ₅ O ₃ S (M+H- t Bu) ⁺ m/z = 470.0; Actual value 470.0.

Step 3. tert-Butyl 4-(7-bromo-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy )-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate

tert -Butyl 4-(7-bromo-6-fluoro-8-methyl-4-(methylsulfinyl)-1 H -[1,2,3]triazolo[4,5) in THF (16 mL) - c ]quinolin-1-yl)piperidine-1-carboxylate (2.51 g, 4.7 mmol) in solution of ( S )-1-(( S )-1-methylpyrrolidin-2-yl)ethane. -1-ol (1.23 g, 9.54 mmol) and DBU (1.44 mL, 9.54 mmol) were added. The mixture was stirred at room temperature overnight, then diluted with EtOAc and extracted with saturated NH ₄ Cl. The combined organic layers were dried over MgSO ₄ , filtered, concentrated and then purified by column chromatography (0-8% MeOH/DCM). LCMS calculated for C ₂₇ H ₃₇ BrFN ₆ O ₃ (M+H) ⁺ m/z = 591.2; Actual value 591.2.

Step 4. 8-(6-Fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1-(piperidine -4-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphtonitrile

Add tert -butyl 4-(7-bromo-6-fluoro-8-methyl-4-(( S )-1-(( S )-1-methylpyrrolidin-2-yl)ethoxy to a reaction vial. )-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate (1.56g, 2.64 mmol), 8-(4,4 ,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-naphtonitrile (1.472 g, 5.27 mmol), SPhos Pd G4 (315 mg, 0.396 mmol), K ₃ PO ₄ (1.679g, 7.91 mmol) and dioxane (11mL) and H ₂ O (2.34mL) were injected. This mixture was sparged with N ₂ for 5 minutes and then heated at 100°C for 3 hours. Upon completion, the reaction was cooled to room temperature, diluted with EtOAc (20 mL), and aq. NH ₄ Cl 20mL). The organic phase was separated, dried over MgSO ₄ , filtered and then concentrated. The concentrated residue was redissolved in CH ₂ Cl ₂ (10 mL) and TFA (5 mL) and stirred at room temperature for 30 minutes. The solvent is then removed and the crude product is purified using preparative-LCMS (XBridge C18 column, eluting with an acetonitrile/water gradient containing 0.1% TFA at a flow rate of 60 mL/min) to give the desired product. Provided as the TFA salt. LCMS calculated for C ₃₃ H ₃₅ FN ₇ O (M+H) ⁺ m/z = 564.3; Actual value 564.2.

Step 5. 8-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrroli din-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphtonitrile

8-(6-fluoro-8-methyl-4-(( S )-1-(( S )-1-methylpyrrolidin-2-yl)ethoxy) in CH ₂ Cl ₂ (0.4 M) -1-(piperidin-4-yl) -1H- [1,2,3]triazolo[4,5- c ]quinolin-7-yl)-1-naphtonitrile solution in triethyl Amine (5 eq) was added followed by a solution of acryloyl chloride (1.5 eq) in CH ₂ Cl ₂ (0.5 M). The mixture was stirred at room temperature for 30 minutes and then the reaction was quenched by addition of MeOH. This mixture was concentrated under reduced pressure, then diluted with AcN and purified by preparative LCMS.

Example 11a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₆ H ₃₇ FN ₇ O ₂ (M+H ⁾ + m/z = 618.3; Actual value 618.2.

Example 11b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₆ H ₃₇ FN ₇ O ₂ (M+H ⁾ + m/z = 618.3; Actual value 618.2. ¹ H NMR (600 MHz, DMSO) δ 9.98 - 9.50 (d, J = 10.7 Hz, 1H), 8.56 - 8.46 (dd, J = 8.5, 1.3 Hz, 1H), 8.38 - 8.24 (m, 2H), 8.16 - 8.10 (dd, J = 7.2, 1.4 Hz, 1H), 7.91 - 7.83 (dd, J = 8.3, 7.0 Hz, 1H), 7.81 - 7.72 (dd, J = 8.3, 7.2 Hz, 1H), 7.71 - 7.64 (d, J = 7.0 Hz, 1H), 6.98 - 6.84 (dd, J = 16.7, 10.5 Hz, 1H), 6.25 - 6.16 (dd, J = 16.8, 2.4 Hz, 1H), 5.85 - 5.77 (m, 1H) ), 5.77 - 5.73 (dd, J = 10.5, 2.4 Hz, 1H), 5.57 - 5.44 (m, 1H), 4.72 - 4.24 (m, 1H), 4.01 - 3.88 (m, 1H), 3.71 - 3.48 (m , 2H), 3.31 - 3.19 (m, 2H), 3.19 - 3.11 (d, J = 4.8 Hz, 3H), 2.48 - 2.34 (m, 2H), 2.34 - 2.03 (m, 9H), 2.00 - 1.86 (m , 2H), 1.64 - 1.52 (d, J = 6.1 Hz, 3H).

Example 12. 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)- 6-fluoro-8-methyl-1H-imidazo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidine-2 -1) Acetonitrile

Step 1. tert-Butyl (2S,4S)-4-((7-bromo-2-chloro-8-fluoro-6-iodo-3-nitroquinolin-4-yl)amino)-2-(cyano Methyl)piperidine-1-carboxylate

DIPEA (4.5 mL, 25.4 mmol) was dissolved in 7-bromo-2,4-dichloro-8-fluoro-6-iodo-3-nitroquinoline ( Intermediate 1 ) (7.9 g, 16.96) in dioxane (50 mL). mmol) and tert -butyl ( 2S , 4S )-4-amino-2-(cyanomethyl)piperidine-1-carboxylate ( Intermediate 7 ) (4.87 g, 20.35 mmol) at room temperature. did. The mixture was stirred at room temperature overnight and then most of the dioxane was removed under reduced pressure. Ice and water were added to the remaining residue, and the slurry was stirred vigorously. The solid precipitate was collected by filtration and dried under air to give the desired product as a yellow solid (quantitative yield). LCMS calculated for C ₂₁ H ₂₂ BrClFIN ₅ O ₄ (M+H) ⁺ : m/z = 668.0; Actual value: 667.8.

Step 2. tert-Butyl (2S,4S)-4-((7-bromo-2-(3-(dimethylamino)-3-methylazetidin-1-yl)-8-fluoro-6-iodo- 3-nitroquinolin-4-yl)amino)-2-(cyanomethyl)piperidine-1-carboxylate

tert -butyl (2 S ,4 S )-4-((7-bromo-2-chloro-8-fluoro-6-iodo-3-nitroquinolin-4-yl)amino in dioxane (20 mL) )-2-(Cyanomethyl)piperidine-1-carboxylate (2.5 g, 3.74 mmol) in a solution of DIPEA (2.61 mL, 14.95 mmol) and N,N ,3-trimethylazetidin-3-amine. Dihydrochloride (1.05 g, 5.61 mmol) was added. The mixture was stirred at 50° C. overnight and then most of the dioxane was removed under reduced pressure. Ice and water were added to the remaining residue, and the slurry was stirred vigorously until a fine powder formed. The solid precipitate was collected through filtration and dried under air to give the desired product (2.7 g, 97% yield). LCMS calculated for C ₂₇ H ₃₅ BrFIN ₇ O ₄ (M+H) ⁺ : m/z = 746.1; Actual value: 746.1.

Step 3. tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-iodo-1H -imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate

tert -Butyl (2 S ,4 S )-4-((7-bromo-2-(3-(dimethylamino)-3-methylazetidin-1-yl)-8-fluoro-6-io A mixture of 3-nitroquinolin-4-yl)amino)-2-(cyanomethyl)piperidine-1-carboxylate (1.0 g, 1.34 mmol) and iron (450 mg, 8.04 mmol) was incubated with AcOH ( 5.0 mL) and heated at 60°C until the nitro group was completely reduced. The mixture was cooled to room temperature, then diluted with EtOAc and filtered over a pad of Celite. The filtrate was concentrated to give tert -butyl (2 S ,4 S )-4-((3-amino-7-bromo-2-(3-(dimethylamino)-3-methylazetidin-1-yl) -8-Fluoro-6-iodoquinolin-4-yl)amino)-2-(cyanomethyl)piperidine-1-carboxylate was obtained. LCMS calculated for C ₂₇ H ₃₇ BrFIN ₇ O ₂ (M+H) ⁺ : m/z = 716.1; Actual value: 716.1.

Triethyl orthoformate (0.45 mL, 2.68 mmol) and toluene (10 mL) were added to the concentrated residue. This mixture was heated at 100° C. overnight, then cooled to room temperature and concentrated. The residue was purified by column chromatography (0-10% MeOH/CH ₂ Cl ₂ ) to obtain the desired product (534 mg, 55% yield). LCMS calculated for C ₂₈ H ₃₅ BrFIN ₇ O ₂ (M+H) ⁺ : m/z = 726.1; Actual value: 726.1.

Step 4. tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H- imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate

This compound is tert -butyl (2 S ,4 S )-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1 H -[1,2,3] triazolo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate with tert -butyl (2 S ,4 S )-4-(7-bro parent-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-iodo- 1H -imidazo[4,5- c ]quinoline-1- Replaced with 1)-2-(cyanomethyl)piperidine-1-carboxylate, prepared according to the procedure described in Intermediate 8, Step 2 . LCMS calculated for C ₂₉ H ₃₈ BrFN ₇ O ₂ (M+H) ⁺ m/z = 614.2; Actual value 614.3.

Step 5. tert-Butyl (2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro -8-methyl-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate

tert -butyl (2 S ,4 S )-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidine-1- in dioxane (5.5 mL)/water (1.1 mL) 1)-6-fluoro-8-methyl- 1H -imidazo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate (785mg, 1.277 mmol), (2-chloro-3-methylphenyl)boronic acid (283 mg, 1.661 mmol), sodium carbonate (542 mg, 5.11 mmol) and Pd(Ph ₃ P) ₄ (221 mg, 0.192 mmol) were evacuated and purified again with nitrogen. Charged (this process was repeated 3 times). The reaction was stirred at 90°C for 3 hours. This mixture was diluted with ethyl acetate and washed with water and brine. The organics were filtered, dried over MgSO ₄ , concentrated and purified by preparative LCMS. LCMS calculated for C ₃₆ H ₄₄ ClFN ₇ O ₂ (M+H) ⁺ m/z = 660.3; Actual value 660.3.

Step 6. 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro -8-methyl-1H-imidazo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)aceto nitrile

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6- Fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1- 1) Piperidine-1-carboxylate is tert -butyl (2 S , 4 S )-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3- Methylazetidin-1-yl)-6-fluoro-8-methyl-1 H -imidazo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl)piperidine-1- Replaced with carboxylate and prepared according to the procedure described in Example 1a and Example 1b , Step 2 . The product was isolated as a mixture of diastereomers. LCMS calculated for C ₃₅ H ₃₉ ClF ₂ N ₇ O (M+H) ⁺ m/z = 646.3; Actual value 646.3. ¹ H NMR (600 MHz, DMSO) δ 10.66 - 10.31 (s, 1H), 8.54 - 8.50 (m, 1H), 7.96 - 7.82 (d, J = 4.8 Hz, 1H), 7.51 - 7.46 (m, 1H) , 7.44 - 7.37 (td, J = 7.6, 2.4 Hz, 1H), 7.29 - 7.17 (d, J = 7.6 Hz, 1H), 6.89 - 6.80 (m, 2H), 5.60 - 5.39 (m, 1H), 5.36 - 4.85 (m, 1H), 5.24 - 5.11 (dd, J = 46.5, 2.5 Hz, 2H), 4.79 - 4.19 (m, 5H), 3.72 - 3.56 (m, 1H), 3.49 - 3.14 (m, 2H) , 2.85 - 2.75 (s, 6H), 2.49 - 2.03 (m, 10H), 1.71 - 1.61 (s, 3H).

Example 13. 2-((2 S ,4 S )-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-(( S )-One-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -Imidazo[4,5- c ]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile

Step 1. tert-Butyl (2S,4S)-4-(7-bromo-4-chloro-6-fluoro-8-iodo-1H-imidazo[4,5-c]quinolin-1-yl)-2 -(Cyanomethyl)piperidine-1-carboxylate

This compound was prepared following the procedure described in Example 12 , Step 1 followed by Step 3 . LCMS calculated for C ₂₂ H ₂₂ BrClFIN ₅ O ₂ (M+H) ⁺ m/z = 648.0; Actual value 648.0.

Step 2. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-imidazo[4,5-c]quinolin-1-yl )-2-(Cyanomethyl)piperidine-1-carboxylate

This compound is tert -butyl (2 S ,4 S )-4-(7-bromo-4-chloro-6-fluoro-8-iodo-1 H -[1,2,3]triazole [4,5- c ]quinolin-1-yl)-2-(2-( tert -butoxy)-2-oxoethyl)piperidine-1-carboxylate with tert -butyl ( 2S , 4S ) -4-(7-bromo-4-chloro-6-fluoro-8-iodo- 1H -imidazo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl)p Replaced with peridine-1-carboxylate, prepared according to the procedure described in Intermediate 4 , Step 2 . LCMS calculated for C ₂₃ H ₂₅ BrFIN ₅ O ₂ S (M+H) ⁺ : m/z = 660.0; Actual value: 660.0.

Step 3. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1H-imidazo[4,5-c]quinolin-1-yl) -2-(Cyanomethyl)piperidine-1-carboxylate

This compound is tert -butyl (2 S ,4 S )-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1 H -[1,2,3] triazolo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate with tert -butyl (2 S ,4 S )-4-(7-bro parent-6-fluoro-8-iodo-4-(methylthio)-1 H -imidazo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl)piperidine-1 -Carboxylate was replaced and prepared according to the procedure described in Intermediate 8 Step 2 . LCMS calculated for C ₂₄ H ₂₈ BrFN ₅ O ₂ S (M+H) ⁺ m/z = 548.1; Actual value 548.1.

Step 4. tert-Butyl (2S,4S)-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-(methylthio)-1H-imidazo[4,5-c ]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate

This compound is tert -butyl ( 2S , 4S )-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro- 8-methyl- 1H -imidazo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate with tert -butyl ( 2S , 4S )- 4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1 H -imidazo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl) Replaced with piperidine-1-carboxylate, prepared according to the procedure described in Example 12, Step 5 . LCMS calculated for C ₃₁ H ₃₄ ClFN ₅ O ₂ S (M+H) ⁺ m/z = 594.2; Actual value 594.3.

Step 5. tert-Butyl (2S,4S)-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methyl Pyrrolidin-2-yl)ethoxy)-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl- 1H- indazole-6- 1)-4-(methylthio) -1H- [1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate was reacted with tert -butyl (2 S ,4 S )-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-(methylthio)-1 H -imidazo[4,5- c ]quinoline -1-yl)-2-(cyanomethyl)piperidine-1-carboxylate was prepared according to the procedure described in Example 7, steps 2 to 3 . LCMS calculated for C ₃₇ H ₄₅ ClFN ₆ O ₃ (M+H) ⁺ m/z = 675.3; Actual value 675.3.

Step 6. 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methyl Pyrrolidin-2-yl)ethoxy)-1H-imidazo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)aceto nitrile

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6- Fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1- 1) Piperidine-1-carboxylate is tert -butyl (2 S ,4 S )-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-(( S )-1-(( S )-1-methylpyrrolidin-2-yl)ethoxy) -1H -imidazo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl )piperidine-1-carboxylate, and replacing ( E )-4-fluorobut-2-enoic acid with 2-fluoroacrylic acid, as described in Examples 1a and 1b , step 2. It was prepared according to the procedure. The product was isolated as a mixture of diastereomers. LCMS calculated for C ₃₅ H ₃₈ ClF ₂ N ₆ O ₂ (M+H) ⁺ m/z = 647.3; Actual value 647.3.

Examples 14a and 14b. 8-(1-((2 R ,4 S )-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-(( S )-One-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]Quinoline-7-yl)-1-naphtonitrile

Step 1. tert-Butyl (2R,4S)-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-[1,2,3]triazolo[ 4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate

This compound is obtained by combining tert -butyl ( 2S , 4S )-4-amino-2-(2-( tert -butoxy)-2-oxoethyl)piperidine-1-carboxylate with tert- butyl (2 R ,4 S )-4-amino-2-methylpiperidine-1-carboxylate was replaced and prepared according to the procedure described in Intermediate 4 . LCMS calculated for C ₂₁ H ₂₅ BrFIN ₅ O ₂ S (M+H) ⁺ : m/z = 636.0; Actual value: 636.0.

Step 2. tert-Butyl (2R,4S)-4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4 ,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate

tert - Butyl ( 2R , 4S )-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio) -1H- [ in dioxane (60mL) and water (20mL) 1,2,3]triazolo[4,5- c ]quinolin-1-yl)-2-methylpiperidine-1-carboxylate (5g, 7.86 mmol), methylboronic acid (0.941g, 15.72 mmol) ), potassium phosphate (5.00 g, 23.57 mmol) and dichlorobis(triphenylphosphine)-palladium(II) (0.827 g, 1.179 mmol) was heated to 90°C and stirred at this temperature for 3 hours. . This mixture was then cooled to room temperature, diluted with AcOEt and water, and separated. The aqueous layer was extracted with AcOEt and the combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and evaporated. The residue was purified by column chromatography (0-20% EtOAc in CH ₂ Cl ₂ ) to provide the desired product as a brown solid. LCMS calculated for C ₂₂ H ₂₈ BrFN ₅ O ₂ S (M+H) ⁺ m/z = 524.1; Actual value 524.1.

Step 3. tert-Butyl (2R,4S)-4-(7-bromo-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidine- 2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl- 1H- indazole-6- 1)-4-(methylthio) -1H- [1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate was reacted with tert -butyl (2 R ,4 S )-4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1 H -[1,2,3]triazolo[4,5- c ] Replaced with quinolin-1-yl)-2-methylpiperidine-1-carboxylate, prepared according to the procedure described in Example 7, steps 2 to 3 . The residue was purified by column chromatography (20-80% AcOEt in DCM) to provide the desired product as a brown solid. LCMS calculated for C ₂₈ H ₃₉ BrFN ₆ O ₃ (M+H) ⁺ m/z = 605.2; Actual value 605.2.

Step 4. tert-Butyl (2R,4S)-4-(7-(8-cyanonaphthalen-1-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1 -methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxyl rate

tert -butyl (2R,4S)-4-(7-bromo-6-fluoro-8-methyl-4-((S)-1-((S)- in dioxane (8 mL) and water (2 mL) 1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidin-1- Carboxylate (200 mg, 0.330 mmol), 8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-naphtonitrile (184 mg, 0.661 mmol) , tripotassium phosphate (351 mg, 1.651 mmol) and SPhos Pd G4 (79 mg, 0.099 mmol) were heated to 90°C. After stirring at the same temperature for 3 hours, the mixture was cooled to room temperature, diluted with EtOAc and water, and separated. The aqueous layer was extracted with AcOEt and the combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and evaporated. The residue was purified by column chromatography (20-80% EtOAc in CH ₂ Cl ₂ ) to provide the desired product as a brown solid. LCMS calculated for C ₃₉ H ₄₅ FN ₇ O ₃ (M+H) ⁺ m/z = 678.4; Actual value 678.4.

Step 5. 8-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1 -((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naph tonitrile

tert -Butyl ( 2R , 4S )-4-(7-(8-cyanonaphthalen-1-yl)-6-fluoro-8-methyl-4-(( S )-1-(( S ) -1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-2-methylpiperidine- 1-Carboxylate (400 mg, 0.578 mmol) was dissolved in 5 mL of TFA. The mixture was stirred at room temperature for 10 minutes and then the solvent was removed.

The residue was dissolved in acetonitrile (10 mL), and Et ₃ N (484 μl, 3.47 mmol) was added. After stirring at 0°C for a few minutes, the cloudy mixture became a clear solution. Acryloyl chloride (94 μl, 1.157 mmol) was added. The mixture was stirred at 0°C for 30 min, then diluted with TFA and preparative-LCMS (XBridge C18 column, eluting with a gradient of acetonitrile/water containing 0.1% TFA at a flow rate of 60 mL/min). This was purified to provide the desired product.

Example 14a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₇ H ₃₉ FN ₇ O ₂ (M+H ⁾ + m/z = 632.3; Actual value 632.3.

Example 14b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₇ H ₃₉ FN ₇ O ₂ (M+H ⁾ + m/z = 632.3; Actual value 632.3.

^1H NMR (500 MHz, DMSO) δ 9.87 (s, 1H), 8.51 (dd, J = 8.5, 1.4 Hz, 1H), 8.31 (dd, J = 8.3, 1.3 Hz, 1H), 8.21 (s, 1H) ), 8.14 (dd, J = 7.2, 1.3 Hz, 1H), 7.87 (dd, J = 8.3, 7.1 Hz, 1H), 7.77 (dd, J = 8.3, 7.2 Hz, 1H), 7.68 (d, J = 7.0 Hz, 1H), 6.93 - 6.87 (m, 1H), 6.16 (dd, J = 16.7, 2.4 Hz, 1H), 5.95 (s, 1H), 5.73 (dd, J = 10.5, 2.4 Hz, 1H), 5.51 (dd, J = 10.1, 5.9 Hz, 1H), 5.08 (s, 1H), 4.68 (s, 1H), 4.27 (d, J = 13.9 Hz, 1H), 3.95 (dd, J = 7.8, 7.7 Hz) , 1H), 3.68 (s, 1H), 3.57 (dq, J = 12.2, 6.2 Hz, 1H), 3.17 (m, J = 4.9 Hz, 3H), 2.46 (d, J = 9.7 Hz, 2H), 2.35 - 2.26 (m, 2H), 2.22 (s, 3H), 2.10 (td, J = 11.8, 5.2 Hz, 1H), 1.93 (tq, J = 14.3, 6.8 Hz, 2H), 1.58 (d, J = 6.1 Hz, 4H), 1.51 (d, J = 7.0 Hz, 2H).

Examples 15a and 15b. 2-((2 S ,4 S )-4-(7-(5,6-dimethyl-1 H -indazol-4-yl)-4-(3-(ethyl(methyl)amino)azetidin-1-yl)-6-fluoro-8-methyl-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile

Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H- indazol-4-yl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)p Peridine-1-carboxylate

Intermediate 8 (1 g, 1.755 mmol), 5,6-dimethyl-1-(tetrahydro- 2H -pyran-2-yl)-4-(4,4,5) in dioxane (12 mL) and water (6 mL) ,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1 H -indazole (0.938 g, 2.63 mmol), tripotassium phosphate (1.862 g, 8.77 mmol) and Pd(Ph ₃ P ) A mixture of ₄ (0.203 g, 0.175 mmol) was heated to 100°C and stirred at this temperature for 2 hours. This mixture was then cooled to room temperature, diluted with AcOEt and water, and separated. The aqueous layer was extracted with EtOAc and the combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and evaporated. The residue was purified by column chromatography (10-40% EtOAc in CH ₂ Cl ₂ ) to provide the desired product as a brown solid. LCMS calculated for C ₃₇ H ₄₄ FN ₈ O ₃ S (M+H) ⁺ m/z = 699.3; Actual value 699.3.

Step 2. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H- indazol-4-yl)-4-(3-(ethyl(methyl)amino)azetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[ 4,5-c]quinolin-1-yl)piperidine-1-carboxylate

This compound is reacted in step 2 with tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4 -(methylthio)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate with tert -butyl (2 S ,4 S )-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro- 2H -pyran-2-yl) -1H -indazol-4-yl)- 6-fluoro-8-methyl-4-(methylthio)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate and replacing N , N ,3-trimethylazetidin-3-amine in step 3 with N -ethyl- N -methylazetidin-3-amine dihydrochloride, Example 6 , steps 2 to 3 It was prepared according to the procedure described in. The crude residue was purified by column chromatography (20-80% EtOAc in CH ₂ Cl ₂ ) to provide the desired product as a brown solid. LCMS calculated for C ₄₂ H ₅₄ FN ₁₀ O ₃ (M+H) ⁺ m/z = 765.4; Actual value 765.3.

Step 3. 2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)azetidine- 1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl ) piperidin-2-yl) acetonitrile

tert- Butyl (2 S ,4 S )-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2 H -pyran-2-yl)-1 H -indazol-4-yl)-4-(3-(ethyl(methyl)amino)azetidin-1-yl)-6-fluoro-8-methyl-1 H -[1,2,3]triazole Ro[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate (378 mg, 0.494 mmol) was dissolved in TFA (5 mL). The mixture was stirred at 60° C. for 10 minutes and then the solvent was removed. The residue was combined with 2-fluoroacrylic acid (131 mg, 1.455 mmol), followed by the addition of acetonitrile (10 mL) followed by DIPEA (506 μl, 2.89 mmol) and T3P (853 μl, 50% in EtOAc, 1.447 mmol). . After stirring overnight at room temperature, the mixture was diluted with TFA, filtered, and preparative-LCMS (XBridge C18 column, eluting with an acetonitrile/water gradient containing 0.1% TFA at a flow rate of 60 mL/min). Purification provided the desired product as the TFA salt.

Example 15a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₅ H ₃₉ F ₂ N ₁₀ O (M+H) ⁺ m/z = 653.3; Actual value 653.3. ^1H NMR (600 MHz, DMSO) δ 13.02 (s, br, 1H), 10.21 (s, br, 1H), 8.09 (s, 1H), 7.47 (s, 1H), 7.30 (s, 1H), 5.86 (td, J = 11.4, 5.5 Hz, 1H), 5.45 - 5.38 (m, 1H), 5.33 (m, 1H), 4.69 (s, 8H), 4.40 (d, J = 8.8 Hz, 1H), 4.15 ( s, 1H), 3.73 (s, 1H), 3.36 (dd, J = 17.2, 6.6 Hz, 2H), 3.11 (m, 1H), 2.86 (s, 3H), 2.49 (d, J = 9.5 Hz, 1H) ), 2.47 (s, 3H), 2.13 (s, 3H), 2.02 (s, 3H), 1.25 (t, J = 7.3 Hz, 3H).

Example 15b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₅ H ₃₉ F ₂ N ₁₀ O (M+H) ⁺ m/z = 653.3; Actual value 653.3.

Example 16a and Example 16b. 8-(6-fluoro-1-(1-(( E )-4-fluorobut-2-enoyl)piperidin-4-yl)-8-methyl-4-(( S )-One-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]Quinoline-7-yl)-1-naphtonitrile

8-(6-fluoro-8-methyl-4-(( S )-1-(( S )-1-methylpyrrolidin-2-yl from Example 11 , Step 1-4) in a round bottom flask. ) Ethoxy)-1-(piperidin-4-yl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)-1-naphtonitrile (600 mg, 0.854 mmol), anhydrous propylphosphonic acid solution (50% in EtOAc, 1.6 mL, 2.56 mmol), ( E )-4-fluorobut-2-enoic acid (267 mg, 2.56 mmol) and acetonitrile ( 15mL) was injected. The reaction flask was cooled to 0°C, and TEA (1.7 mL, 12.8 mmol) was added to the reaction. After stirring at room temperature for 2 hours, the reaction was purified with aq. The reaction was stopped with NaHCO ₃ (10 mL), extracted with EtOAc (25 mL×3), and concentrated. The crude mixture was redissolved in acetonitrile and purified using preparative-LCMS (XBridge C18 column, eluting with an acetonitrile/water gradient containing 0.1% TFA at a flow rate of 60 mL/min) to yield the desired product. provided.

Example 16a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₇ H ₃₈ F ₂ N ₇ O ₂ (M+H) ⁺ : m/z = 650.3; Actual value: 650.3.

Example 16b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₇ H ₃₈ F ₂ N ₇ O ₂ (M+H) ⁺ : m/z = 650.3; Actual value: 650.3.

Example 17a and Example 17b. 8-(1-((2 S ,4 S )-2-(cyanomethyl)-1-(2-fluoroacryloyl)piperidin-4-yl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl )-6-fluoro-8-methyl-1 H -[1,2,3]triazolo[4,5- c ]Quinoline-7-yl)-1-naphtonitrile

Step 1. tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H- [1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methyl Replace thio)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate with intermediate 8 , Example 6 , Step 2 It was prepared according to the procedures described in 3 to 3 . The crude residue was purified on silica (0-10% MeOH in DCM) to provide the desired product as a brown solid (700 mg, 63% yield). LCMS calculated for C ₂₈ H ₃₇ BrFN ₈ O ₂ (M+H) ⁺ : m/z = 615.2; Actual value: 615.2.

Step 2. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(7-(8-cyanonaphthalen-1-yl)-4-(3-(dimethylamino)-3- methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxyl rate

Add tert -butyl ( 2S , 4S )-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8 to the reaction vial. -Methyl-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate (500 mg, 0.79 mmol ), SPhos Pd G4 (97 mg, 0.12 mmol), 8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-naphtonitrile (453 mg, 1.63 mmol), K ₃ PO ₄ (571 mg, 2.44 mmol), and dioxane (9 mL) and H ₂ O (3 mL) were injected. This mixture was sparged with N ₂ for 5 minutes and then heated at 90°C for 2 hours. Upon completion, the reaction was cooled to room temperature, diluted with EtOAc (30 mL), and aq. Washed with water (20mL). The organic phase was separated, dried over MgSO ₄ and purified on silica (0-10% MeOH in DCM) to give the desired product as a brown solid (358 mg, 64% yield). LCMS calculated for C ₃₉ H ₄₃ FN ₉ O ₂ (M+H) ⁺ : m/z = 688.4; Actual value: 688.3.

Step 3. 8-(1-((2S,4S)-2-(cyanomethyl)-1-(2-fluoroacryloyl)piperidin-4-yl)-4-(3-(di Methylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)- 1-Naphtonitrile

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6- Fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1- 1) Piperidine-1-carboxylate is tert -butyl (2 S ,4 S )-2-(cyanomethyl)-4-(7-(8-cyanonaphthalen-1-yl)-4-( 3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl- 1H -[1,2,3]triazolo[4,5- c ]quinoline- 1-day) Replace with piperidine-1-carboxylate and ( E )-4-Fluorobut-2-enoic acid was replaced with 2-fluoroacrylic acid, prepared according to the procedure described in Example 1a and Example 1b , Step 2 .

Example 17a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₇ H ₃₆ F ₂ N ₉ O (M+H) ⁺ : m/z = 660.3; Actual value: 660.3. ^1H NMR (600 MHz, DMSO) δ 8.48 (dd, J = 8.4, 1.3 Hz, 1H), 8.28 (dd, J = 8.4, 1.3 Hz, 1H), 8.12 (dd, J = 7.1, 1.3 Hz, 1H) ), 8.06 (s, 1H), 7.85 (dd, J = 8.3, 7.1 Hz, 1H), 7.75 (dd, J = 8.3, 7.1 Hz, 1H), 7.62 (dd, J = 7.1, 1.3 Hz, 1H) , 5.90-5.75 (m, 1H), 5.45-5.30 (m, 2H), 4.00-4.50 (m, 11H), 2.85 (s, 6H), 2.50 (s, 1H), 2.20-2.15 (m, 4H) , 1.70 (s, 3H).

Example 17b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₇ H ₃₆ F ₂ N ₉ O (M+H) ⁺ : m/z = 660.3; Actual value: 660.3.

Examples 18a and 18b. 2-((2 S ,4 S )-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoroquinolin-8-yl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1-yl)-1-(( E )-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile

Step 1. 7-Bromo-6,8-dichloro-2H-benzo[d][1,3]oxazine-2,4(1H)-dione

NCS (5.6 g, 41.9 mmol) was added to a solution of 2-amino-4-bromo-3-chlorobenzoic acid (10.0 g, 39.9 mmol) in 1,4-dioxane (100 mL) and the reaction was then incubated at 70°C. It was stirred overnight. After the mixture was cooled to room temperature, triphosgene (4.7 g, 15.96 mmol) was added, and then stirred at 80° C. for 2 hours. Upon completion, ice was added and the mixture was stirred for 30 minutes. The solid precipitate was collected via filtration and washed with ice water to provide the desired product.

Step 2. 7-Bromo-6,8-dichloro-3-nitroquinoline-2,4-diol

DIPEA (3.49 mL, 19.97 mmol) was added to a solution of ethyl 2-nitroacetate (2.66 g, 19.97 mmol) in toluene (40.0 mL) at room temperature. After stirring for 10 minutes, 7-bromo-6,8-dichloro- 2H -benzo[ d ][1,3]oxazine-2,4( 1H )-dione (3.1g, 10.0 mmol) was added, and the resulting mixture was stirred at 95°C for 3 hours. The reaction was cooled with ice water and the solid was then collected via filtration and washed with a small amount of ethyl acetate to provide the desired product as a yellow solid (1.05 g, 30%). LCMS calculated for C ₉ H ₄ BrCl ₂ N ₂ O ₄ (M+H) ⁺ : m/z = 352.9; Actual value 353.0.

Step 3. 7-Bromo-2,4,6,8-tetrachloro-3-nitroquinoline

DIPEA (0.98 mL, 5.6 mmol) was dissolved in 7-bromo-6,8-dichloro-3-nitroquinoline-2,4-diol (1.0 g, 2.82 mmol) in POCl ₃ (6.0 mL, 64.4 mmol). was added to the mixture at room temperature. The reaction was then stirred at 100°C for 3 hours. The solvent was removed under vacuum. The concentrated residue was redissolved in ethyl acetate and then washed with NaHCO ₃ (aq.). The combined organic layers were concentrated under vacuum to give the crude product, which was used directly in the next step without further purification.

Step 4. tert-Butyl (2S,4S)-4-((7-bromo-6,8-dichloro-2-(3-(dimethylamino)-3-methylazetidin-1-yl)- 3-nitroquinolin-4-yl)amino)-2-(cyanomethyl)piperidine-1-carboxylate

In a solution of 7-bromo-2,4,6,8-tetrachloro-3-nitroquinoline (1 g, 2.56 mmol) in CH ₂ Cl ₂ (10 mL) DIPEA (1.8 mL, 10.24 mmol) and intermediate 7 730 mg, 3.0 mmol) was added. The mixture was stirred at room temperature overnight, then N,N,3-trimethylazetidin-3-amine (350 mg, 3.0 mmol) was added and the mixture was stirred at room temperature until completion. The reaction was diluted with CH ₂ Cl ₂ and then washed with saturated NH ₄ Cl solution. The solvent was removed under vacuum and the crude product was used directly in the next step without further purification. LCMS calculated for C ₂₇ H ₃₅ BrCl ₂ N ₇ O ₄ (M+H) ⁺ : m/z = 670.1; Actual value 670.1.

Step 5. tert-Butyl (2S,4S)-4-(7-bromo-6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-1H -[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate

tert -butyl (2 S ,4 S )-4-((7-bromo-6,8-dichloro-2-(3-) in THF/MeOH/H ₂ O (1:1:1) (30 mL) (dimethylamino)-3-methylazetidin-1-yl)-3-nitroquinolin-4-yl)amino)-2-(cyanomethyl)piperidine-1-carboxylate (1.7g, 2.56 mmol), iron (1.43 g, 25.6 mmol) and NH ₄ Cl (1.37 g, 25.6 mmol) were heated at 65°C for 30 min. Upon completion, the reaction was diluted with MeOH and filtered. The solvent was removed under vacuum and the residue was basified with NaHCO ₃ and extracted with CH ₂ Cl ₂ . The solvent was removed under vacuum.

The concentrated residue was re-dissolved in acetic acid (5 mL), and sodium nitrite (176 mg, 5.12 mmol) was added. This mixture was stirred at room temperature for 1 hour. On completion, the solvent was removed under vacuum and the residue was basified with NaHCO ₃ and extracted with CH ₂ Cl ₂ . The combined organic layers were dried over MgSO4, filtered, concentrated to dryness, and purified on silica gel to give the desired product (1.0 g, 60%). LCMS calculated for C ₂₇ H ₃₄ BrCl ₂ N ₈ O ₂ (M+H) ⁺ : m/z = 651.1; Actual value 651.0.

Step 6. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl )-7-(5-fluoroquinolin-8-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate

tert -butyl (2 S ,4 S )-4-(7-bromo-6,8-dichloro-4-(3-(dimethylamino)- in dioxane/H ₂ O = 4:1 (10 mL) 3-methylazetidin-1-yl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-2-(cyanomethyl)piperidine-1 -In a solution of carboxylate (650 mg, 1.0 mmol), K ₃ PO ₄ (850 mg, 4 mmol), (5-fluoroquinolin-8-yl)boronic acid (286 mg, 1.5 mmol) and Pd(PPh ₃ ) ₄ (80 mg, 0.07 mmol) was added and the reaction mixture was heated to 90° C. for 1.5 hours. Upon completion, the reaction was diluted with water, extracted with CH ₂ Cl ₂ and the combined organic layers were dried over MgSO ₄ , filtered, concentrated to dryness and purified on silica gel to give the desired product as a mixture of diastereomers. (550 mg, 76%). LCMS calculated for C ₃₆ H ₃₉ Cl ₂ FN ₉ O ₂ (M+H) ⁺ : m/z = 718.3; Actual value 718.2.

Step 7. 2-((2S,4S)-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluo Roquinolin-8-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-ene Oil) piperidin-2-yl) acetonitrile

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7- (2,3-dimethylphenyl)-6-fluoro-8-methyl-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1- The carboxylate was reacted with tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidine-1 -yl)-7-(5-fluoroquinolin-8-yl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1- Replaced with carboxylate and prepared according to the procedure described in Example 6 , Step 4 . The product was purified using pre-LCMS (XBridge C18 column, flow rate of 60 mL/min, eluting with a gradient of acetonitrile/water containing 0.15% NH ₄ OH) to provide the desired product.

Example 18a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₆ H ₃₇ Cl ₂ FN ₉ O ₂ (M+H) ⁺ m/z = 716.2; Actual value 716.2.

Example 18b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₆ H ₃₇ Cl ₂ FN ₉ O ₂ (M+H) ⁺ m/z = 716.2; Actual value 716.2.

Examples 19a and 19b. 2-((2 S ,4 S )-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoroquinolin-8-yl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1-yl)-1-(( E )-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile

This compound is tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6- Fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1- I) piperidine-1-carboxylate tert -butyl (2 S ,4 S )-2-(cyanomethyl)-4-(6,8-dichloro-4-(3-(dimethylamino) -3-methylazetidin-1-yl)-7-(5-fluoroquinolin-8-yl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1- 1) Replaced with piperidine-1-carboxylate, prepared according to the procedure described in Example 1a and Example 1b , Step 2 .

Example 19a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₅ H ₃₄ Cl ₂ F ₂ N ₉ O (M+H) ⁺ m/z = 704.2; Actual value 704.2.

Example 19b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₅ H ₃₄ Cl ₂ F ₂ N ₉ O (M+H) ⁺ m/z = 704.2; Actual value 704.2.

Example 20. 2-((2 S ,4 S )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(tri fluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1-yl)-1-(( E )-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile

This compound is similar to Example 1a and Example 1b by replacing ( E )-4-fluorobut-2-enoic acid with (E) -4-methoxybut-2-enoic acid. Prepared according to the procedure described in Step 2 .

Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro- 7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperi Din-1-carboxylate

Intermediate 3 (1.3g, 1.9 mmol), XPhos Pd G2 (76mg, 0.097 mmol), (5-fluoroquinolin-8-yl)boronic acid (408mg, 2.1 mmol), K ₃ PO ₄ (1.24g) in a reaction vial. , 5.83 mmol) and dioxane (5 mL) and H ₂ O (1 mL) were injected. This mixture was sparged with N ₂ for 5 minutes and then heated at 90°C for 1 hour. Upon completion, the reaction was cooled to room temperature, diluted with EtOAc (20 mL), and aq. Washed with NH ₄ Cl (20 mL). The organic phase was separated, dried over MgSO ₄ , filtered and then concentrated. The crude product was first purified by silica (20 g, 50-100% EtOAc in CH ₂ Cl ₂ ) and then purified by preparative-LCMS (XBridge C18 column, flow rate of 60 mL/min, containing 0.15% NH ₄ OH). Further purification using an acetonitrile/water gradient) separated the diastereomers (white amorphous powder, 21% combined yield).

Diastereomer 1 . Peak 1. LCMS calculated for C ₃₇ H ₃₉ F ₅ N ₉ O ₂ (M+H) ⁺ m/z = 736.3; Actual value 736.2.

Diastereomer 2 . Peak 2. LCMS calculated for C ₃₇ H ₃₉ F ₅ N ₉ O ₂ (M+H) ⁺ m/z = 736.3; Actual value 736.2.

Step 2. 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinoline -8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4- Methoxybut-2-enoyl)piperidin-2-yl)acetonitrile

tert -butyl (2 S ,4 S )-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6- from Step 1 Fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinoline-1- 1) TFA (1 mL) was added to the reaction vial injected with piperidine-1-carboxylate ( diastereomer 1 , 300 mg, 0.41 mmol) at room temperature. After stirring for 15 minutes, volatile substances were removed. The residue was redissolved in acetonitrile (4 mL) and cooled to 0°C. To this reaction was added DIPEA (0.21 mL), followed by ( E )-4-methoxybut-2-enoic acid (71 mg, 0.61 mmol) and anhydrous propylphosphonic acid solution (50% in EtOAc, 0.50 mL, 0.82 mmol). Added. After stirring at 0°C for 10 min, the reaction was incubated with aq. The reaction was quenched with NaHCO ₃ (5 mL), extracted with EtOAc (5 mL), washed with brine and concentrated. The crude product was dissolved in acetonitrile and purified using preparative-LCMS (XBridge C18 column, eluting with a gradient of acetonitrile/water containing 0.1% TFA at a flow rate of 60 mL/min) to give Example 20 ( single diastereomer) was provided as the TFA salt in the form of a white amorphous powder. LCMS calculation for C ₃₇ H ₃₇ F ₅ N ₉ O ₂ (M+H) ⁺ m/z = 734.3; Actual value 734.4. ^1H NMR (600 MHz, DMSO-d6) δ 10.80 (s, 1H), 8.87 (dd, J = 4.2, 1.7 Hz, 1H), 8.62 (dd, J = 8.5, 1.8 Hz, 1H), 8.41 (s) , 1H), 7.82 (dd, J = 8.0, 5.9 Hz, 1H), 7.70 (dd, J = 8.5, 4.1 Hz, 1H), 7.65 (dd, J = 9.7, 8.0 Hz, 1H), 6.81 - 6.72 ( m, 2H), 5.93 (s, 1H), 5.31 (s, 1H), 4.96 (m, 2H), 4.76 - 4.32 (m, 2H), 4.11 (d, J = 3.1 Hz, 2H), 3.67 - 3.34 (m, 2H), 3.33 (s, 3H), 3.19 (td, J = 16.6, 10.1 Hz, 2H), 2.84 (s, 6H), 2.49 - 2.43 (m, 2H), 2.33 (s, 1H), 2.21 (d, J = 15.7 Hz, 1H), 1.71 (s, 3H).

Example 21. 2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-( (S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E )-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile

Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-4-((S)-1-((S)-1-methylp rolidin-2-yl)ethoxy)-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-[1,2,3]tri Azolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate

Add tert-butyl (2S,4S)-4-(7-bromo-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidine- 2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate ( 315 mg, 0.5 mmol), bis(pinacolato)diborone (165 mg, 0.65 mmol), Pd(dppf)Cl ₂ (36 mg, 0.05 mmol), potassium acetate (147 mg, 1.5 mmol), and dioxane (5 mL) were injected. . This mixture was sparged with N ₂ for 5 minutes and then heated at 80° C. for 16 hours. Upon completion, the reaction was cooled to room temperature, diluted with DCM (100 mL), and aq. Washed with NH ₄ Cl (20 mL). The organic phase was separated, dried over MgSO ₄ , filtered and then concentrated. The crude product was used in the next step without further purification.

Step 2. 2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-(( S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl ) Acetonitrile

Crude tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-4-((S)-1-(( S)-1-methylpyrrolidin-2-yl)ethoxy)-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-[ 1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate, 1-chloro-4-fluoroisoquinoline (118mg, 0.65 mmol), Pd( PPh ₃ ) ₄ (58 mg, 0.05 mmol), K ₃ PO ₄ (318 mg, 1.5 mmol), dioxane (4 mL), and water (1 mL) were injected. This mixture was heated at 90°C for 1 hour. Upon completion, the reaction was cooled to room temperature, diluted with ethyl acetate (100 mL), and aq. Washed with NH ₄ Cl (20 mL). The organic phase was separated, dried over MgSO ₄ , filtered and then concentrated.

The crude product was dissolved in 5 mL DCM/TFA (1:1) solution. Upon completion, the reaction was concentrated. The crude product was redissolved in acetonitrile and purified using preparative-LCMS (XBridge C18 column, eluting with an acetonitrile/water gradient containing 0.1% TFA at a flow rate of 60 mL/min) to give the desired product. was given as a mixture of diastereomers, LCMS calculated m/z for C ₃₃ H ₃₅ F ₂ N ₈ O (M+H) ⁺ = 597.3; Actual value 597.3.

Step 3. 2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-(( S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E) -4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile

2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-(( S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl ) Acetonitrile (30 mg, 0.05 mmol), DIPEA (0.087 mL, 0.5 mmol), ( E )-4-methoxybut-2-enoic acid (12 mg, 0.1 mmol), 1 mL MeCN and anhydrous propylphosphonic acid solution (50% in EtOAc, 0.06 mL, 0.1 mmol) was injected. After stirring at 0°C for 10 min, the reaction was diluted with acetonitrile and preparative-LCMS (XBridge C18 column, eluting with an acetonitrile/water gradient containing 0.1% TFA at a flow rate of 60 mL/min). Purification gave the desired product as a mixture of diastereomers.

Example 21 . LCMS calculation for C ₃₈ H ₄₁ F ₂ N ₈ O ₃ (M+H) ⁺ m/z = 695.3; Actual value 695.3.

Example 22. 2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-( (S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2- Fluoroacryloyl)piperidin-2-yl)acetonitrile

This compound was prepared following the procedure described in Example 21 , replacing ( E )-4-methoxybut-2-enoic acid with 2-fluoroacrylic acid.

Example 22 . LCMS calculation for C ₃₆ H ₃₆ F ₃ N ₈ O ₂ (M+H) ⁺ m/z = 669.3; Actual value 669.3.

Example 23a and Example 23b. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1 -methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl ) piperidin-2-yl) acetonitrile

Step 1. 3-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquinoline

4-Bromo-3-methylisoquinoline (1.110 g, 5.0 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2 in dioxane (20.0 mL) A mixture of '-bi(1,3,2-dioxaborolane) (2.285 g, 9.00 mmol), potassium acetate (1.472 g, 15.00 mmol) and PdCl ₂ (dppf) (0.366 g, 0.500 mmol) was incubated at 105°C. It was stirred for 5 hours. The solvent was removed and the product was purified by column eluting with hexane/EtOAc (up to 80% EtOAc). LCMS calculated for C ₁₆ H ₂₁ BNO ₂ (M+H) ⁺ m/z = 270.1; Actual value 270.1.

Step 2. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S )-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl) Acetonitrile

tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-8-methyl-4-((S)-1-((S)) in dioxane (2 mL)/water (0.4 mL) -1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)p Peridine-1-carboxylate (45.0 mg, 0.071 mmol), 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquinoline ( 28.8 mg, 0.107 mmol), potassium phosphate (45.4 mg, 0.214 mmol) and methanesulfonato (2-dicyclohexylphosphino-2',6'-dimethoxy-1,1'-biphenyl) (2' A mixture of -methylamino-1,1'-biphenyl-2-yl)palladium(II) dichloromethane adduct (SPhos Pd G4) (5.68 mg, 7.14 μmol) was vacuumed and then recharged twice with N2. Then, the reaction was stirred at 95°C for 4 hours. This mixture was diluted with MeCN and then purified by preparative-HPLC at PH = 10. The solvent was removed and the residue was dissolved in MeCN (1.0 mL), then TFA (1.0 mL) was added and the reaction was stirred at room temperature for 40 min. The solvent was removed and the crude product was used directly in the next step. LCMS calculated for C ₃₄ H ₃₈ FN ₈ O (M+H) ⁺ m/z = 593.3; Actual value 593.4.

Step 3. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S )-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoro Acryloyl)piperidin-2-yl)acetonitrile

Triethylamine (5.64 μl, 0.040 mmol) was dissolved in 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinoline-4-) in ethyl acetate (0.8 mL). 1)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c ]quinolin-1-yl)piperidin-2-yl)acetonitrile (4.0 mg, 6.75 μmol), 2-fluoroacrylic acid (1.215 mg, 0.013 mmol) and 1-propanephosphonic acid cyclic anhydride (T3P in EtOAc) 50%) (4.02 μl, 0.013 mmol) was added at 0° C., and the reaction was then stirred for 30 min. The solvent was removed and the crude product was redissolved in acetonitrile and preparative-LCMS (XBridge C18 column, eluting with a gradient of acetonitrile/water containing 0.1% TFA at a flow rate of 60 mL/min). Purification provided the product as the TFA salt.

Example 23a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₇ H ₃₉ F ₂ N ₈ O ₂ (M+H) ⁺ m/z = 665.3; Actual value 665.4.

Example 23b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₇ H ₃₉ F ₂ N ₈ O ₂ (M+H) ⁺ m/z = 665.3; Actual value 665.4. ¹ H NMR (600 MHz, DMSO) δ 9.46 (s, 1H), 8.32 (s, 1H), 8.27 (m, 1H), 7.72 (m, 2H), 7.22 (m, 1H), 5.94 (m 1H) , 5.64 (m, 1H), 5.28-5.37 (m, 2H), 3.96 (q, 1H), 3.63 (m, 3H), 3.54 (m, 1H), 3.38 (m, 2H), 3.21 (m, 1H) ), 3.16 (s, 3H), 2.51 (m, 4H), 2.40 (s, 3H), 2.34 (m, 1H), 2.19 (s, 3H), 2.13 (m, 1H), 1.97 (m, 2H) , 1.59 (d, 3H).

Example 24a and Example 24b. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1 -methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-meth Toxybut-2-enoyl)piperidin-2-yl)acetonitrile

This compound was prepared following the procedure described in Examples 23a and 23b , Step 3, replacing 2-fluoroacrylic acid with (E)-4-methoxybut- 2-enoic acid.

Example 24a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₉ H ₄₄ FN ₈ O ₃ (M+H ⁾ + m/z = 691.3; Actual value 691.5.

Example 24b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₉ H ₄₄ FN ₈ O ₃ (M+H ⁾ + m/z = 691.3; Actual value 691.5.

Example 25a and Example 25b. 2-((2S,4S)-4-(6-fluoro-7-(7-fluoro-2-methylquinolin-8-yl)-8-methyl-4-((S)-1-(( S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluo Roacryloyl) piperidin-2-yl) acetonitrile

Step 1. 2-((2S,4S)-4-(6-fluoro-7-(7-fluoro-2-methylquinolin-8-yl)-8-methyl-4-((S)-1 -((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine- 2-day) Acetonitrile

This compound is 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquinoline (7-fluoro-2-methylquinoline- 8-day) replaced with boronic acid and prepared according to the procedure described in Example 23a and Example 23b , Step 2 . LCMS calculated for C ₃₄ H ₃₇ F ₂ N ₈ O (M+H) ⁺ m/z = 611.3; Actual value 611.4.

Step 2. 2-((2S,4S)-4-(6-fluoro-7-(7-fluoro-2-methylquinolin-8-yl)-8-methyl-4-((S)-1 -((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-( 2-fluoroacryloyl)piperidin-2-yl)acetonitrile

This compound is 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-(( S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl ) Acetonitrile is 2-((2S,4S)-4-(6-fluoro-7-(7-fluoro-2-methylquinolin-8-yl)-8-methyl-4-((S) -1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperi din-2-yl)acetonitrile, prepared according to the procedure described in Example 23a and Example 23b , Step 3 .

Example 25a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₇ H ₃₈ F ₃ N ₈ O ₂ (M+H) ⁺ m/z = 683.3; Actual value 683.4.

Example 25b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₇ H ₃₈ F ₃ N ₈ O ₂ (M+H) ⁺ m/z = 683.3; Actual value 683.4.

Example 26a and Example 26b. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methylisoquinolin-4-yl)-4-((S)-1-((S)-1 -methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-meth Toxybut-2-enoyl)piperidin-2-yl)acetonitrile

Step 1. (1-methylisoquinolin-4-yl)boronic acid

4-Bromo-1-methylisoquinoline (0.222 g, 1.0 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2 in dioxane (5.0 mL) A mixture of '-bi(1,3,2-dioxaborolane) (0.457 g, 1.800 mmol), potassium acetate (0.294 g, 3.00 mmol) and PdCl2(dppf) (0.073 g, 0.100 mmol) was incubated at 105°C for 4 hours. Stirred for an hour. The product was purified by preparative-HPLC at PH = 2 (TFA). LCMS calculated for C ₁₀ H ₁₁ BNO ₂ (M+H) ⁺ m/z = 188.1; Actual value 188.1.

Step 2. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methylisoquinolin-4-yl)-4-((S)-1-((S )-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl) Acetonitrile

This compound is 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquinoline (1-methylisoquinolin-4-yl) Replaced with boronic acid and prepared according to the procedure described in Example 23a and Example 23b , Step 2 . LCMS calculated for C ₃₄ H ₃₈ FN ₈ O (M+H) ⁺ m/z = 593.3; Actual value 593.4.

Step 3. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methylisoquinolin-4-yl)-4-((S)-1-((S )-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)- 4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile

Triethylamine (5.64 μl, 0.040 mmol) was dissolved in 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methylisoquinoline-4-) in ethyl acetate (0.8 mL). 1)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c ]quinolin-1-yl)piperidin-2-yl)acetonitrile (4.0 mg, 6.75 μmol), (E)-4-methoxybut-2-enoic acid (1.567 mg, 0.013 mmol) and 1- A solution of propanephosphonic acid cyclic anhydride (T3P 50% in EtOAc) (4.02 μl, 0.013 mmol) was added at 0° C. and the reaction was then stirred for 30 min. The solvent was removed and the crude product was redissolved in acetonitrile and preparative-LCMS (XBridge C18 column, eluting with a gradient of acetonitrile/water containing 0.1% TFA at a flow rate of 60 mL/min). Purification provided the product as the TFA salt.

Example 26a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₉ H ₄₄ FN ₈ O ₃ (M+H ⁾ + m/z = 691.4; Actual value 691.5.

Example 26b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₉ H ₄₄ FN ₈ O ₃ (M+H ⁾ + m/z = 691.4; Actual value 691.5.

Example 27a and Example 27b. 2-((2S,4S)-4-(6-fluoro-7-(7-fluoroquinolin-8-yl)-8-methyl-4-((S)-1-((S)-1 -methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl ) piperidin-2-yl) acetonitrile

Step 1. 2-((2S,4S)-4-(6-fluoro-7-(7-fluoroquinolin-8-yl)-8-methyl-4-((S)-1-((S )-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl) Acetonitrile

This compound is 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquinoline (7-fluoroquinolin-8-yl) Replaced with boronic acid and prepared according to the procedure described in Example 23a and Example 23b , Step 2 . LCMS calculated for C ₃₃ H ₃₅ F ₂ N ₈ O (M+H) ⁺ m/z = 597.3; Actual value 597.4.

Step 2. 2-((2S,4S)-4-(6-fluoro-7-(7-fluoroquinolin-8-yl)-8-methyl-4-((S)-1-((S )-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoro Acryloyl)piperidin-2-yl)acetonitrile

This compound is 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-(( S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl ) Acetonitrile is 2-((2S,4S)-4-(6-fluoro-7-(7-fluoroquinolin-8-yl)-8-methyl-4-((S)-1-( (S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2- 1) Replaced with acetonitrile and prepared according to the procedure described in Example 23a and Example 23b , Step 3 .

Example 27a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₆ H ₃₆ F ₃ N ₈ O ₂ (M+H) ⁺ m/z = 669.3; Actual value 669.4.

Example 27b . Diastereomer 2. Peak 2. . LCMS calculation for C ₃₆ H ₃₆ F ₃ N ₈ O ₂ (M+H) ⁺ m/z = 669.3; Actual value 669.4.

Example 28a and Example 28b. 2-(4-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1 -((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl -1H-indazol-3-yl)acetonitrile

Step 1. (4-Bromo-1-methyl-1H-indazol-3-yl)methanol

Sodium hydride (0.34 g, 8.47 mmol; 60% dispersion in mineral oil) was added at 0°C. The reaction mixture was stirred for 5 minutes and allowed to warm to room temperature, at which point it was stirred overnight. Upon completion, the reaction mixture was aq. The reaction was stopped with NH ₄ Cl (5 mL). The aqueous phase was extracted with EtOAc (10 mL×3), dried over MgSO ₄ , filtered and concentrated. The crude mixture was then dissolved in THF (20 mL) and cooled to -78°C, at which point LiAlH ₄ (11.7 mL, 11.7 mmol, 1M in THF) was added slowly by syringe. The reaction mixture was warmed to 0° C. and stirred for a further 15 minutes, at which point aq. NH ₄ Cl (10 mL) was added slowly. The aqueous phase was extracted with EtOAc (10 mL×3), dried over MgSO ₄ , filtered and concentrated. The crude mixture was used in the next step without further purification. LCMS calculated for C ₉ H ₁₀ BrN ₂ O (M+H) ⁺ m/z = 241.0; Actual value 241.0.

Step 2. 2-(4-Bromo-1-methyl-1H-indazol-3-yl)acetonitrile

PBr ₃ (0.39 mL, 4.16 mmol) in (4-bromo-1-methyl-1 H -indazol-3-yl)methanol (0.67 g, 2.77 mmol) in CH ₂ Cl ₂ (10 mL) cooled at -78°C. mmol) was added slowly. The reaction mixture was stirred at the same temperature for 1 hour, at which point the reaction mixture was stirred with aq. The reaction was carefully stopped with NaHCO ₃ (5 mL). The aqueous phase was extracted with EtOAc (20 mL×3), dried over MgSO ₄ , filtered and concentrated. To the crude mixture was added DMSO (8 mL) followed by NaCN (0.41 g, 8.32 mmol) at room temperature. The reaction mixture was stirred at 60° C. for 3 hours and returned to room temperature upon completion. Water (20 mL) was added to the reaction flask, and the desired product was pulverized. The desired product was then collected by filtration, washed with water and air-dried for 4 hours. LCMS calculated for C ₁₀ H ₉ BrN ₃ (M+H) ⁺ m/z = 250.0; Actual value 250.0.

Step 3. 2-(1-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazol-3-yl)acetonite reel

2-(4-bromo-1-methyl-1 H -indazol-3-yl)acetonitrile (0.12 g, 0.48 mmol) in dioxane (3 mL), 4,4,4',4',5, 5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (0.18 g, 0.72 mmol), potassium acetate (0.15 g, 1.54 mmol) and PdCl ₂ ( dppf) (0.024g, 0.033 mmol) was stirred at 95°C for 3 hours. The solvent was removed and the product was purified by silica gel column eluting with hexanes/EtOAc (maximum EtOAc 50%). LCMS calculated for C ₁₆ H ₂₁ BN ₃ O ₂ (M+H) ⁺ m/z = 298.2; Actual value 298.2.

Step 4. tert-Butyl (2R,4S)-4-(7-(3-(cyanomethyl)-1-methyl-1H-indazol-4-yl)-6-fluoro-8-methyl-4-(( S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl) -2-methylpiperidine-1-carboxylate

tert -butyl (2 R,4 S )-4-(7-bromo-6-fluoro-8 from Examples 14a and 14b , Step 3 in dioxane/H ₂ O = 4:1 (2 mL) -methyl-4-(( S )-1-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-2-methylpiperidine-1-carboxylate (40 mg, 0.066 mmol) in a solution of K ₃ PO ₄ (49 mg, 0.23 mmol), 2-(1-methyl-4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1 H -indazol-3-yl)acetonitrile (29 mg, 0.099 mmol) and SPhosPdG4 (7.9 mg, 0.01 mmol) was added and the reaction mixture was heated to 90° C. for 1.5 hours. Upon completion, the reaction was diluted with water, extracted with EtOAc (2 mL×3), dried over MgSO ₄ , filtered, and concentrated. The crude mixture was purified on silica gel eluting with CH ₂ Cl ₂ /MeOH (max. MeOH 10%) to give the desired product as a mixture of diastereomers (20 mg, 44%). LCMS calculated for C ₃₈ H ₄₇ FN ₉ O ₃ (M+H) ⁺ : m/z = 696.4; Actual value 696.4.

Step 5. 2-(4-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1 -((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl -1H-indazol-3-yl)acetonitrile

This compound is tert -butyl (2 R ,4 S )-4-(7-(8-cyanonaphthalen-1-yl)-6-fluoro-8-methyl-4-(( S )-1- (( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-2-methyl Piperidine-1-carboxylate was reacted with tert -butyl (2 R ,4 S )-4-(7-(3-(cyanomethyl)-1-methyl-1 H -indazol-4-yl)-6 -fluoro-8-methyl-4-(( S )-1-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo [4,5- c ]quinolin-1-yl)-2-methylpiperidine-1-carboxylate, prepared according to Example 14a and Example 14b, Step 5 . This mixture was purified using pre-LCMS (XBridge C18 column, eluting with an acetonitrile/water gradient containing 0.1% TFA at a flow rate of 60 mL/min) to provide the desired product.

Example 28a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₆ H ₄₁ FN ₉ O ₂ (M+H ⁾ + m/z = 650.3; Actual value 650.3.

Example 28b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₆ H ₄₁ FN ₉ O ₂ (M+H ⁾ + m/z = 650.3; Actual value 650.3.

Example 29. 2-((2S,4S)-4-(4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6- Fluoro-5-methyl-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-( 2-fluoroacryloyl)piperidin-2-yl)acetonitrile

Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran- 2-yl)-1H-indazol-4-yl)-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl ) Piperidine-1-carboxylate

Intermediate 8 (2 g, 3.64 mmol) in dioxane (25 mL)/water (12 mL), 6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-4-(4,4, 5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (1.967 g, 5.46 mmol), tripotassium phosphate (3.86 g, 18.20 mmol) and Pd(Ph ₃ P ) The mixture of ₄ (0.421 g, 0.364 mmol) was brought to vacuum and recharged twice with N ₂ and the reaction was then stirred at 102° C. for 2 hours. The mixture was then cooled to room temperature, diluted with AcOEt and water, and separated. The aqueous layer was extracted with EtOAc and the combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and evaporated. The residue was purified by column chromatography (10-40% EtOAc in CH ₂ Cl ₂ ) to provide the desired product as a brown solid. LCMS calculation for C ₃₆ H ₄₁ F ₂ N ₈ O ₃ S (M+H) ⁺ m/z = 703.3; Actual value 703.3.

Step 2. 2-((2S,4S)-4-(4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro Ro-5-methyl-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-2 -1) Acetonitrile

m CPBA (77% wet) (0.842 g, 3.76 mmol) was tert -butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-) in CH ₂ Cl ₂ (30 mL). (6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-methyl-4-(methylthio)-1H-[1 ,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (2.4 g, 3.41 mmol) was added at 0°C, and this reaction was then The temperature was stirred for 20 minutes. The reaction was quenched by adding saturated aqueous Na ₂ S ₂ O ₃ and Na ₂ CO ₃ , diluted with CH ₂ Cl ₂ and separated. The aqueous layer was extracted with CH ₂ Cl ₂ . The combined organic layers were dried over Na ₂ SO ₄ and concentrated to give tert -butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-(6-fluoro-5-methyl- 1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-methyl-4-(methylsulfinyl)-1H-[1,2,3]triazolo[ A mixture of 4,5-c]quinolin-1-yl)piperidine-1-carboxylate and the corresponding sulfonyl compound was provided, which was used directly in the next step.

tert -Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl )-1H-indazol-4-yl)-8-methyl-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)p Combine peridine-1-carboxylate (and the corresponding sulfonyl compound, 2.4 g, 3.34 mmol) and N-ethyl-N,3-dimethylazetidin-3-amine (0.428 g, 3.34 mmol) and acetonite. Reel (33.4 mL) was added. DIPEA (1.166 ml, 6.68 mmol) was added to this suspension and the mixture was then heated to 70° C. and stirred at the same temperature for 2 hours. The solvent was evaporated to give the crude product as a brown solid.

tert -Butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7 -(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]tri Azolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate was dissolved in 10 ml of TFA, heated to 60° C. and stirred at the same temperature for 10 minutes. Then cooled to room temperature, diluted with acetonitrile, filtered, and purified using prep-LCMS (XBridge C18 column, eluting with a gradient of acetonitrile/water containing 0.1% TFA at a flow rate of 60 mL/min). The diastereomers (white amorphous powder, 53% combined yield) were separated.

Diastereomer 1 . Peak 1. LCMS calculated for C ₃₂ H ₃₇ F ₂ N ₁₀ (M+H) ⁺ m/z = 599.3; Actual value 599.3.

Diastereomer 2 . Peak 2. LCMS calculated for C ₃₂ H ₃₇ F ₂ N ₁₀ (M+H) ⁺ m/z = 599.3; Actual value 599.3.

Step 3. 2-((2S,4S)-4-(4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro Ro-5-methyl-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2 -Fluoroacryloyl)piperidin-2-yl)acetonitrile

2-((2S,4S)-4-(4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5 -methyl-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl) Acetonitrile bis(2,2,2-trifluoroacetate) ( diastereomer 2 , 395 mg, 0.477 mmol) was combined with 2-fluoroacrylic acid (134 mg, 1.488 mmol) followed by acetonitrile (10 mL). This was followed by the addition of DIPEA (780 μl, 4.46 mmol) and T3P (877 μl, 50% in AcOEt, 1.488 mmol). After stirring overnight at room temperature, the mixture was diluted with TFA, filtered, and preparative-LCMS (XBridge C18 column, eluting with an acetonitrile/water gradient containing 0.1% TFA at a flow rate of 60 mL/min). and purified to provide Example 29 (single diastereomer) as a TFA salt in the form of a white amorphous powder.

Example 29 . LCMS calculation for C ₃₅ H ₃₈ F ₃ N ₁₀ O (M+H) ⁺ m/z = 671.3; Actual value 671.3. ^1H NMR (600 MHz, DMSO) δ 13.28 (s, br, 1H), 10.16 (s, br, 1H), 8.12 (s, 1H), 7.47-7.44 (m, 2H), 5.86 (td, J = 11.4, 5.5 Hz, 1H), 5.45 - 5.38 (m, 2H), 5.16 (m, 1H), 4.77-4.73 (m, 2H), 4.50-4.36 (m, 4H), 3.66-3.44 (m, 2H) , 3.36 (m, 2H), 3.11 (m, 1H), 2.81 (s, 3H), 2.49-2.45 (m, 2H), 2.17 (s, 3H), 2.03 (s, 3H), 1.71 (s, 3H) ), 1.27 (t, J = 7.3 Hz, 3H).

Example 30. 2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3 -methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2- Fluoroacryloyl)piperidin-2-yl)acetonitrile

Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H- indazol-4-yl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)p Peridine-1-carboxylate

Intermediate 8 (2g, 3.64 mmol), 5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5, in dioxane (25mL)/water (12mL) 5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (1.945 g, 5.46 mmol), tripotassium phosphate (3.86 g, 18.20 mmol) and Pd(Ph ₃ P) ₄ The mixture of (0.421 g, 0.364 mmol) was evacuated and recharged twice with N ₂ and the reaction was then stirred at 102° C. for 2 h. This mixture was then cooled to room temperature, diluted with AcOEt and water, and separated. The aqueous layer was extracted with EtOAc and the combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and evaporated. The residue was purified by column chromatography (10-40% EtOAc in CH ₂ Cl ₂ ) to provide the desired product as a brown solid. LCMS calculated for C ₃₇ H ₄₄ FN ₈ O ₃ S (M+H) ⁺ m/z = 699.3; Actual value 699.3.

Step 2. 2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3- methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl ) Acetonitrile

mCPBA (0.962 g, 77% wet, 4.29 mmol) was tert- butyl ( _2S ,4S)-2-(cyanomethyl)-4-(7-(5,6-di) in CH ₂ Cl 2 (30 mL). Methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2, Added to a solution of 3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (2 g, 2.86 mmol) at 0°C, and then incubated the reaction at this temperature for 20 minutes. It was stirred for a while. The reaction was quenched by adding saturated aqueous Na ₂ S ₂ O ₃ and Na ₂ CO ₃ , diluted with CH ₂ Cl ₂ and separated. The aqueous layer was extracted with CH ₂ Cl ₂ . The combined organic layers were dried over Na ₂ SO ₄ and concentrated to give tert -butyl (2S,4S)-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2H) -pyran-2-yl)-1H-indazol-4-yl)-6-fluoro-8-methyl-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4, A crude mixture of 5-c]quinolin-1-yl)piperidine-1-carboxylate and the corresponding sulfonyl compound was provided, which was used directly in the next step.

tert- Butyl (2S,4S)-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol- 4-yl)-6-fluoro-8-methyl-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine -1-Carboxylate (and the corresponding sulfonyl compound, 1.5 g, 2.098 mmol), N-ethyl-N,3-dimethylazetidin-3-amine (0.323 g, 2.52 mmol) were combined, and acetonitrile ( 20mL) was added. DIPEA (1.832 ml, 10.49 mmol) was added to this suspension and the mixture was then heated to 70° C. and stirred at the same temperature for 2 hours. The solvent was evaporated to give the crude product as a brown solid.

tert- butyl (2S,4S)-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-inda obtained above. zol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]tri Azolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate was dissolved in 10 ml of TFA, heated to 60° C. and stirred at the same temperature for 10 minutes. Then cooled to room temperature, diluted with acetonitrile, filtered, and purified using prep-LCMS (XBridge C18 column, eluting with a gradient of acetonitrile/water containing 0.1% TFA at a flow rate of 60 mL/min). The diastereomers (white amorphous powder, 51% combined yield) were separated.

Diastereomer 1 . Peak 1. LCMS calculated for C ₃₃ H ₄₀ FN ₁₀ (M+H) ⁺ m/z = 595.3; Actual value 595.3.

Diastereomer 2 . Peak 2. LCMS calculated for C ₃₃ H ₄₀ FN ₁₀ (M+H) ⁺ m/z = 595.3; Actual value 595.3.

Step 3. 2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3- Methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoro Roacryloyl) piperidin-2-yl) acetonitrile

2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidine -1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonite Combine lil bis(2,2,2-trifluoroacetate) ( diastereomer 2 , 400 mg, 0.486 mmol) with 2-fluoroacrylic acid (88 mg, 0.972 mmol), followed by acetonitrile (10 mL). DIPEA (849 μl, 4.86 mmol) and T3P (573 μl, 50% in AcOEt, 0.972 mmol) were added. After stirring overnight at room temperature, the mixture was diluted with TFA, filtered, and preparative-LCMS (XBridge C18 column, eluting with an acetonitrile/water gradient containing 0.1% TFA at a flow rate of 60 mL/min). Purification provided Example 30 (single diastereomer) as the TFA salt in the form of a white amorphous powder.

Example 30 . LCMS calculated for C ₃₆ H ₄₁ F ₂ N ₁₀ O (M+H) ⁺ m/z = 667.3; Actual value 667.3. ^1H NMR (600 MHz, DMSO) δ 13.02 (s, br, 1H), 10.18 (s, br, 1H), 8.10 (s, 1H), 7.47 (s, 1H), 7.31 (d, J = 5.1 Hz , 1H), 5.86 (td, J = 11.4, 5.5 Hz, 1H), 5.41 (dd, J = 18.2, 4.4 Hz, 1H), 5.38-4.62 (m, 9H), 3.73-3.56 (m, 2H), 3.36 (dd, J = 16.9, 6.8 Hz, 1H), 3.07 (td, J = 14.0, 5.2 Hz, 1H), 2.81 (s, 3H), 2.49 (s, 3H), 2.48-2.45 (m, 2H) , 2.14 (s, 3H), 2.02 (s, 3H), 1.71 (s, 3H), 1.27 (t, J = 7.3 Hz, 3H).

Example 31a and Example 31b. 2-(4-(1-((2 R ,4 S )-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-(( S )-One-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)-1-methyl-1 H -indole-3-yl)acetonitrile

Step 1. 2-(1-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)acetonitrile

This compound is made by combining 2-(4-bromo-1-methyl-1 H -indazol-3-yl)acetonitrile with 2-(4-bromo-1-methyl-1 H -indole-3-yl) ) Replaced with acetonitrile and prepared according to Example 28a and Example 28b, Step 3 . This mixture was purified using silica gel column chromatography eluting with hexane/EtOAc (maximum EtOAc 50%). LCMS calculated for C ₁₇ H ₂₂ BN ₂ O ₂ (M+H) ⁺ m/z = 297.2; Actual value 297.2.

Step 2. tert-Butyl (2R,4S)-4-(7-(3-(cyanomethyl)-1-methyl-1H-indol-4-yl)-6-fluoro-8-methyl-4-((S )-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)- 2-methylpiperidine-1-carboxylate

This compound is 2-(1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1 H -indazol-3-yl) Acetonitrile was dissolved in 2-(1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1 H -indol-3-yl)aceto. Prepared according to Examples 28a and 28b, Step 4, replacing with nitrile . The crude mixture was purified on silica gel eluting with CH ₂ Cl ₂ /MeOH (maximum MeOH 10%) to give the desired product as a mixture of diastereomers. LCMS calculated for C ₃₉ H ₄₈ FN ₈ O ₃ (M+H) ⁺ : m/z = 695.4; Actual value 695.4.

Step 3. 2-(4-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1 -((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl -1H-indole-3-yl)acetonitrile

This compound is tert -butyl (2 R ,4 S )-4-(7-(8-cyanonaphthalen-1-yl)-6-fluoro-8-methyl-4-(( S )-1- (( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-2-methyl Piperidine-1-carboxylate is reacted with tert -butyl (2 R ,4 S )-4-(7-(3-(cyanomethyl)-1-methyl-1 H -indol-4-yl)-6- Fluoro-8-methyl-4-(( S )-1-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[ 4,5- c ]quinolin-1-yl)-2-methylpiperidine-1-carboxylate, prepared according to Example 14a and Example 14b, Step 5 . This mixture was purified using preparative-LCMS (XBridge C18 column, eluting with an acetonitrile/water gradient containing 0.1% TFA at a flow rate of 60 mL/min) to provide the desired product.

Example 31a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₇ H ₄₂ FN ₈ O ₂ (M+H ⁾ + m/z = 649.3; Actual value 649.3.

Example 31b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₇ H ₄₂ FN ₈ O ₂ (M+H ⁾ + m/z = 649.3; Actual value 649.3. ^1H NMR (600 MHz, DMSO) δ 8.17 (s, 1H), 7.62 (d, J = 8.3 Hz, 1H), 7.48 (s, 1H), 7.37 (dd, J = 8.3, 7.1 Hz, 1H), 7.00 (d, J = 7.1 Hz, 1H), 6.96-6.84 (m, 1H), 6.16 (dd, J = 16.6, 2.4 Hz, 1H), 5.95-5.85 (m, 1H), 5.73 (dd, J = 10.5, 2.4 Hz, 1H), 5.57 (dq, J = 12.4, 6.2 Hz, 1H), 4.73-4.62 (m, 1H), 4.31-4.20 (m, 1H), 3.97-3.91 (m, 1H), 3.88 (s, 3H), 3.65-3.54 (m, 2H), 3.20-3.07 (m, 6H), 2.51-2.45 (m, 2H), 2.38-2.25 (m, 2H), 2.24 (s, 3H), 2.15 -1.90 (m, 4H), 1.57-1.43 (m, 6H).

Example 32a and Example 32b. 2-(4-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-(( S )-One-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)-1-methyl-1 H -indole-3-yl)acetonitrile

Step 1. tert-Butyl 4-(7-(3-(cyanomethyl)-1-methyl-1H-indol-4-yl)-6-fluoro-8-methyl-4-((S)-1-(( S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxyl rate

This compound was prepared by reacting 8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-naphtonitrile in Examples 31a and 31b , Step 1. from 2-(1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1 H -indol-3-yl)acetonitrile Alternately, prepared according to Example 11a and Example 11b, Step 4 . The crude mixture was purified on silica gel eluting with CH ₂ Cl ₂ /MeOH (maximum MeOH 10%) to give the desired product as a mixture of diastereomers. LCMS calculated for C ₃₈ H ₄₆ FN ₈ O ₃ (M+H) ⁺ m/z = 681.4; Actual value 681.4.

Step 2. 2-(4-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrroli din-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl-1H-indol-3-yl)acetonite reel

This compound is tert -butyl (2 R ,4 S )-4-(7-(8-cyanonaphthalen-1-yl)-6-fluoro-8-methyl-4-(( S )-1- (( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-2-methyl Piperidine-1-carboxylate was reacted with tert -butyl 4-(7-(3-(cyanomethyl)-1-methyl-1 H -indol-4-yl)-6-fluoro-8-methyl-4 -(( S )-1-(( S )-1-methylpyrrolidin-2-yl)ethoxy) -1H- [1,2,3]triazolo[4,5- c ]quinoline- 1-yl)piperidine-1-carboxylate, prepared according to Example 14a and Example 14b, Step 5 . This mixture was purified using pre-LCMS (XBridge C18 column, eluting with an acetonitrile/water gradient containing 0.1% TFA at a flow rate of 60 mL/min) to provide the desired product.

Example 32a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₆ H ₄₀ FN ₈ O ₂ (M+H ⁾ + m/z = 635.3; Actual value 635.3.

Example 32b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₆ H ₄₀ FN ₈ O ₂ (M+H ⁾ + m/z = 635.3; Actual value 635.3. ¹ H NMR (600 MHz, DMSO) δ 8.26 (s, 1H), 7.61 (d, J = 8.4 Hz, 1H), 7.47 (s, 1H), 7.45-7.35 (m, 1H), 7.00 (d, J = 7.1 Hz, 1H), 6.93 (dd, J = 16.7, 104 Hz, 1H), 6.18 (dd, J = 16.7, 2.5 Hz, 1H), 5.79-5.70 (m, 2H), 5.59-5.50 (m, 1H), 4.62-4.53 (m, 1H), 4.31-4.24 (m, 1H), 3.99-3.90 (m, 1H), 3.87 (s, 3H), 3.70-3.11 (m, 10H), 2.51-2.18 ( m, 8H), 1.58 (d, J = 6.1 Hz, 3H).

Examples 33a and 33b. 2-((2S,4S)-4-(8-chloro-7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3 -methylazetidin-1-yl)-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacrylo 1) piperidine-2-yl) acetonitrile

Step 1. 7-Bromo-2,4,6-trichloro-8-fluoro-3-nitroquinoline

This compound was prepared following the procedure described in Intermediate 1, replacing NIS with NCS in Step 1 . LCMS calculated for C ₉ H ₂ BrCl ₃ FN ₂ O ₂ (M+H) ⁺ m/z = 372.8; Actual value 372.8.

Step 2. tert-Butyl (2S,4S)-4-(7-bromo-8-chloro-6-fluoro-4-(methylthio)-1H-[1,2,3]triazolo[4 ,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate

This compound is obtained by combining 7-bromo-2,4-dichloro-8-fluoro-6-iodo-3-nitroquinoline with 7-bromo-2,4,6-trichloro-8-fluoro Replaced with -3-nitroquinoline, prepared according to the procedure described in Intermediate 8 , Step 1 . LCMS calculated for C ₂₂ H ₂₄ BrClFN ₆ O ₂ S (M+H) ⁺ m/z = 569.1; Actual value 569.1.

Step 3. tert-Butyl (2S,4S)-4-(8-chloro-7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4 -yl)-6-fluoro-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)p Peridine-1-carboxylate

tert -butyl (2S,4S)-4-(7-bromo-8-chloro-6-fluoro-4-(methylthio)-1H-[1, in dioxane (11.70 mL)/water (5.85 mL) 2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate (1g, 1.755 mmol), 5,6-dimethyl- 1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (0.938 g, 2.63 mmol), tripotassium phosphate (1.862 g, 8.77 mmol) and Pd(Ph ₃ P) ₄ (0.203 g, 0.175 mmol) were evacuated and recharged twice with N ₂ and then the reaction was It was stirred at 102°C for 2 hours. The mixture was then cooled to room temperature, diluted with AcOEt and water, and separated. The aqueous layer was extracted with EtOAc and the combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and evaporated. The residue was purified by column chromatography (10-40% EtOAc in CH ₂ Cl ₂ ) to provide the desired product as a brown solid. LCMS calculated for C ₃₆ H ₄₁ ClFN ₈ O ₃ S (M+H) ⁺ m/z = 719.3; Actual value 719.3.

Step 4. tert-Butyl (2S,4S)-4-(8-chloro-7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl) -4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinoline -1-yl)-2-(cyanomethyl)piperidine-1-carboxylate

This compound is reacted in step 2 with tert -butyl ( 2S , 4S )-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4- (methylthio) -1H- [1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidine-1-carboxylate with tert- butyl (2S,4S)-4 -(8-chloro-7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-6-fluoro-4-(methyl Replace with thio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate and Step 3 Prepared according to the procedure described in Example 6 , steps 2-3, replacing N , N , 3-trimethylazetidin-3-amine with N-ethyl-N, 3-dimethylazetidin-3-amine. did. The crude residue was purified by column chromatography (20-80% EtOAc in CH ₂ Cl ₂ ) to provide the desired product as a brown solid. LCMS calculated for C ₄₂ H ₅₃ ClFN ₁₀ O ₃ (M+H) ⁺ m/z = 799.4; Actual value 799.4.

Step 5. 2-((2S,4S)-4-(8-chloro-7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino )-3-methylazetidin-1-yl)-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoro Roacryloyl) piperidin-2-yl) acetonitrile

tert-Butyl (2S,4S)-4-(8-chloro-7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl) -4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinoline -1-yl)-2-(cyanomethyl)piperidine-1-carboxylate (427 mg, 0.534 mmol) was dissolved in TFA (5 mL). The mixture was stirred at 60° C. for 10 minutes and then the solvent was removed.

The residue was combined with 2-fluoroacrylic acid (144 mg, 1.601 mmol) followed by the addition of acetonitrile (10 mL) followed by DIPEA (0.93 mL, 5.34 mmol) and T3P (0.94 mL, 50% in AcOEt, 1.601 mmol). did. After stirring overnight at room temperature, the mixture was diluted with TFA, filtered, and preparative-LCMS (XBridge C18 column, eluting with an acetonitrile/water gradient containing 0.1% TFA at a flow rate of 60 mL/min). Purification provided the desired product as the TFA salt.

Example 33a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₅ H ₃₈ ClF ₂ N ₁₀ O (M+H) ⁺ m/z = 687.3; Actual value 687.3.

Example 33b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₅ H ₃₈ ClF ₂ N ₁₀ O (M+H) ⁺ m/z = 687.3; Actual value 687.3.

Example 34a and Example 34b. 2-((2S,4S)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-8-methyl-4-((S)-1-((S)- 1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4- Methoxybut-2-enoyl)piperidin-2-yl)acetonitrile

Step 1. 2-((2S,4S)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-8-methyl-4-((S)-1-(( S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl ) Acetonitrile

This compound is 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquinoline (2-fluoro-6-methoxyphenyl) ) was replaced with boronic acid and prepared according to the procedure described in Example 23a and Example 23b , Step 2 . LCMS calculated for C ₃₁ H ₃₆ F ₂ N ₇ O ₂ (M+H) ⁺ m/z = 576.3; Actual value 576.4.

Step 2. 2-((2S,4S)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-8-methyl-4-((S)-1-(( S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E) -4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile

This compound is 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methylisoquinolin-4-yl)-4-((S)-1-(( S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl ) Acetonitrile is 2-((2S,4S)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-8-methyl-4-((S)-1- ((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2 -1) Replaced with acetonitrile and prepared according to the procedure described in Example 26a and Example 26b , Step 3 .

Example 34a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₆ H ₄₂ F ₂ N ₇ O ₄ (M+H) ⁺ m/z = 674.3; Actual value 674.3.

Example 34b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₆ H ₄₂ F ₂ N ₇ O ₄ (M+H) ⁺ m/z = 674.3; Actual value 674.3.

Examples 35a and 35b. 2-((2S,4S)-4-(8-chloro-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6- Fluoro-5-methyl-1H-indazol-4-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoro Acryloyl)piperidin-2-yl)acetonitrile

This compound is 5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioc in step 3 saborolan-2-yl)-1H-indazole was reacted with 6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl -1,3,2-dioxaborolan-2-yl)-1H-indazole was prepared according to the procedure described in Example 33 .

Example 35a . Diastereomer 1. Peak 1. LCMS calculated for C ₃₄ H ₃₅ ClF ₃ N ₁₀ O (M+H) ⁺ m/z = 691.3; Actual value 691.3.

Example 35b . Diastereomer 2. Peak 2. LCMS calculated for C ₃₄ H ₃₅ ClF ₃ N ₁₀ O (M+H) ⁺ m/z = 691.3; Actual value 691.3.

Example A. GDP-GTP exchange assay.

The inhibitory potency of the exemplified compounds was determined in a fluorescence-based guanine nucleotide exchange assay, which measures the exchange of bodipy-GDP (a fluorescently labeled GDP) for GppNHp (a non-hydrolyzable GTP analogue) in SOS1 ( produces an active state of KRAS in the presence of guanine nucleotide exchange factor. Inhibitors were serially diluted in DMSO and 0.1 μL volumes were transferred to wells of a black low volume 384-well plate. A 5 μL/well volume of bodipy-loaded KRAS G12C diluted to 5 nM in assay buffer (25mM Hepes pH 7.5, 50mM NaCl, 10mM MgCl2, and 0.01% Brij-35) was added to the plate and incubated with inhibitors. Pre-incubated together at ambient temperature for 2 hours. Appropriate controls (enzyme without inhibitor or with G12C inhibitor (AMG-510)) were included in the plate. Exchange was initiated by addition of a 5 μL/well volume containing 1mM GppNHp and 300nM SOS1 in assay buffer. Bodifi-loaded KRAS G12C, GppNHp and SOS1 at 10 μL/well reaction concentrations were 2.5 nM, 500 uM and 150 nM, respectively. Reaction plates were incubated at ambient temperature for 2 hours, the time assessed for complete GDP-GTP exchange in the absence of inhibitors. For KRAS G12D and G12V mutants, a similar guanine nucleotide exchange assay was performed with 2.5 nM as a final concentration for bodifi loaded KRAS protein and 4 and 3 h incubation after addition of the GppNHp-SOS1 mixture for G12D and G12V, respectively. It was used as. The G12D mutant (Sakamoto et al., BBRC 484.3 (2017), 605-611) or a cyclic peptide described as binding selectively to an internal compound with confirmed binding was used as a positive control in the assay plate. Fluorescence intensity was measured on a PheraStar plate reader device (BMG Labtech) with excitation at 485 nm and emission at 520 nm.

Data were analyzed using GraphPad Prism or Gene Data Screener SmartFit. IC ₅₀ values were derived by fitting the data to a four parameter logistic equation that generates a sigmoidal dose-response curve using the variable Hill coefficient.

KRAS_G12C Exchange Assay IC ₅₀ data, KRAS_G12C pERK Assay IC ₅₀ data, KRAS_G12C WB pERK Assay IC ₅₀ data are provided in Table 1 below. The symbol “†” indicates IC ₅₀ ≤50nM, and “††” indicates IC ₅₀ greater than 50nM but less than or equal to 100nM; And “†††” indicates that the IC ₅₀ is greater than 100 nM but less than 1000 nM, and “††††” indicates that the IC ₅₀ is greater than 1 μM but less than 5 μM; “†††††” indicates an IC ₅₀ greater than 5 μM, and “NA” indicates that data is not available.

Example B: Luminescent viability assay

MIA PaCa-2 (KRAS G12C; ATCC® CRL-1420), NCI-H358 (KRAS G12C; ATCC® CRL-5807), A427 (KRAS G12D; ATCC® HTB53), HPAFII (KRAS G12D; ATCC® CRL-1997) , YAPC (KRAS G12V; DSMZ ACC382), SW480 (KRAS G12V; ATCC® CRL-228), and NCI-H838 (KRAS WT; ATCC® CRL-5844) cells were cultured in RPMI 1640 medium supplemented with 10% FBS (Gibco/Life Technologies). White, clean bottom containing 50 nL dots of test compound (final concentration is 1:500 dilution with final concentration in 0.2% DMSO), 800 cells per well in RPMI 1640 medium supplemented with 2% FBS. Seed in 384-well Costar tissue culture plates. Plates are incubated at 37°C, 5% CO2 for 3 days. At the end of the assay, 25 ul/well of CellTiter-Glo reagent (Promega) is added. Luminescence is read after 15 minutes using PHERAstar (BMG). Data are analyzed in Genedata Screener using SmartFit for IC ₅₀ values.

Example C: Cellular pERK HTRF Assay

MIA PaCa-2 (KRAS G12C; ATCC® CRL-1420), NCI-H358 (KRAS G12C; ATCC® CRL-5807), A427 (KRAS G12D; ATCC® HTB53), HPAFII (KRAS G12D; ATCC® CRL-1997) , YAPC (KRAS G12V; DSMZ ACC382), SW480 (KRAS G12V; ATCC® CRL-228), and NCI-H838 (KRAS WT; ATCC® CRL-5844) cells were purchased from ATCC and cultured in RPMI 1640 supplemented with 10% FBS. (Gibco/Life Technologies) medium. Cells are plated at 5000 cells per well (8uL) in Greiner 384-well low volume, flat-bottom tissue culture treated white plates and incubated overnight at 37°C, 5% CO2. The next morning, test compound stock solutions are diluted in medium to a final concentration of 3× and 4 uL are added to the cells to a final concentration of 0.1% in DMSO. Cells are incubated with test compounds for 4 hours (G12C and G12V) or 2 hours (G12D) at 37°C, 5% CO2. 4 uL of 4× lysis buffer with blocking reagent (Cisbio) is added to each well and the plate is rotated gently (300 rpm) for 30 minutes at room temperature. Mix 4uL of Cisbio anti-Phospho-ERK 1/2 d2 per well with anti-Phospho-ERK 1/2 Cryptate (1:1), add to each well, and incubate overnight at room temperature in the dark. Plates are read at 665 nm and 620 nm wavelengths on a Pherastar plate reader. Data are analyzed in Genedata Screener using SmartFit for IC ₅₀ values.

Example D: Whole Blood pERK1/2 HTRF Assay

MIA PaCa-2 cells (KRAS G12C; ATCC® CRL-1420), HPAF-II (KRAS G12D; ATCC® CRL-1997) and YAPC (KRAS G12V; DSMZ ACC382) were cultured in RPMI 1640 containing 10% FBS (Gibco/ Life Technologies). For the MIA PaCa-2 assay, cells were seeded in 96-well tissue culture plates (Corning #3596) at 25,000 cells per well in 100uL medium and cultured for 2 days at 37°C, 5% CO ₂ before assay. For HPAF-II and YAPC assays, cells are seeded in 96 well tissue culture plates at 50000 cells per well in 100 uL medium and cultured for 1 day before assay. Whole blood is added to a 96 well plate in a 1 uL dot of compound (prepared in DMSO) and mixed gently by pipetting up and down to bring the concentration of compound in the blood to the desired concentration of 1× in 0.5% DMSO. Media was aspirated from the cells, and 50 uL of whole blood per well was added with test compounds and incubated at 37°C, 5% CO ₂ for 4 hours for MIA PaCa and YAPC assays, respectively; Alternatively, incubate for 2 hours for HPAF-II assay. After discarding the blood, the plate is washed gently twice by adding PBS to the side of the well, discarding the PBS from the plate onto a paper towel, and taping the plate to ensure good drainage. Then, 50ul/well of 1× Lysis Buffer #1 (Cisbio) containing blocking reagent (Cisbio) and Benzonase nuclease (Sigma Cat # E1014-5KU, 1:10000 final concentration) was added and incubated at room temperature for 30 μl. Incubate with shaking (250 rpm) for 1 minute. After lysis, 16 uL of lysate was transferred to a 384-well Greiner small volume white plate using Assist Plus (Integra Biosciences, NH). A 1:1 mixture of 4 uL of anti-Phospho-ERK 1/2 d2 and anti-Phospho-ERK 1/2 Cryptate (Cisbio) is added to the wells using Assist Plus and incubated overnight at room temperature in the dark. Plates are read at 665 nm and 620 nm wavelengths on a Pherastar plate reader. Data are analyzed in Genedata Screener using SmartFit for IC ₅₀ values.

Example E: Ras Activation Elisa

The 96-well Ras activation ELISA kit (Cell Biolabs Inc; #STA441) selectively knocks down the active form of Ras from cell lysates using the Raf1 Rho binding domain (RBD) bound to 96-well plates. The captured GTP-Ras is then detected by pan-Ras antibody and HRP-conjugated secondary antibody.

MIA PaCa-2 (KRAS G12C; ATCC® CRL-1420), NCI-H358 (KRAS G12C; ATCC® CRL-5807), A427 (KRAS G12D; ATCC® HTB53), HPAFII (KRAS G12D; ATCC® CRL-1997) , YAPC (KRAS G12V; DSMZ ACC382), SW480 (KRAS G12V; ATCC® CRL-228), and NCI-H838 (KRAS WT; ATCC® CRL-5844) cells were cultured in RPMI 1640 (Gibco/Life Technologies) with 10% FBS. ) is maintained. Cells were seeded at 25000 cells per well in 100uL medium in 96 well tissue culture plates (Corning #3596) and cultured for 2 days at 37°C, 5% CO ₂ to approximately 80% confluence at the start of the assay. Cells are treated with compounds for 4 hours or overnight at 37°C, 5% CO ₂ . Upon harvest, cells are washed with PBS, drained well, and then lysed with 50 uL of 1× lysis buffer (provided by the kit) + added Halt protease and phosphatase inhibitors (1:100) for 1 hour on ice.

Raf-1 RBD is diluted 1:500 in assay dilution (provided in the kit) and 100 μL of diluted Raf-1 RBD is added to each well of the Raf-1 RBD capture plate. Cover the plate with plate sealing film and incubate for 1 hour at room temperature on an orbital shaker. Plates are washed three times with 250 μL 1X wash buffer per well with thorough aspiration between each wash. Add 50 μL of Ras lysate sample (10 to 100 μg) to each well in duplicate. A “no cell lysate” control is added to a pair of wells for background determination. 50 μL of assay diluent is added to all wells immediately, and the plate is incubated for 1 hour at room temperature on an orbital shaker. Wash the plate 5 times with 250 μL of 1X wash buffer per well with thorough aspiration between each wash. 100 μL of diluted Anti-pan-Ras Antibody is added to each well, and the plate is incubated for 1 hour on an orbital shaker. Wash the plate five times as before. 100 μL of diluted secondary antibody, HRP conjugate is added to each well and the plate is incubated for 1 hour at room temperature on an orbital shaker. Wash the plate five times as before and drain well. Add 100 mL of Chemiluminescent Reagent (provided in the kit) to each well, including the blank well. The plate is incubated for 5 minutes at room temperature on an orbital shaker and then the luminescence of each microwell is read on the plate emitter. % inhibition is calculated relative to DMSO control wells after subtracting the background level of the “no lysate control” from all values. IC ₅₀ determinations are performed by fitting a curve of inhibitor percent inhibition versus logarithm of inhibitor concentration using GraphPad Prism 7 software.

Example F: Inhibition of RAS-RAF and PI3K-AKT pathways

cells of the compound by measuring phosphorylation of KRAS downstream effectors extracellular-signal-regulated kinase (ERK), ribosomal S6 kinase (RSK), AKT (also known as protein kinase B, PKB) and downstream substrate S6 ribosomal protein. Efficacy was determined.

To measure phosphorylated extracellular-signal-regulated kinase (ERK), ribosomal S6 kinase (RSK), AKT and S6 ribosomal proteins, cells (details regarding type of cell line and data produced are further detailed in Table 2) described) were seeded overnight in Corning 96-well tissue culture treated plates at 4×104 cells/well in RPMI medium containing 10% FBS. The next day, cells were incubated for 4 hours at 37°C, 5% CO ₂ in the presence or absence of a range of concentrations of test compounds. Cells were washed with PBS and lysed with 1× lysis buffer (Cisbio) containing protease and phosphatase inhibitors (Thermo Fisher, 78446). 10 or 20 μg of total protein lysate was subjected to SDS-PAGE and immunoblot analysis using the following antibodies: phospho-ERK1/2-Thr202/Tyr204 (#9101L), total-ERK1/2 (#9102L), phospho-AKT-Ser473 (#4060L), phospho-p90RSK-Ser380 (#11989S), and phospho-S6 ribosomal protein-Ser235/Ser236 (#2211S) were from Cell Signaling Technologies (Danver, MA). ) obtained from

Example G: In Vivo Efficacy Study

Mia-Paca-2 (KRAS G12C), H358 (KRAS G12C), HPAF-II (KRAS G12D), AGS (KRAS G12D), SW480 (KRAS G12V), or YAPC (KRAS G12V) human cancer cells were collected from the American Type Culture Collection. Type Culture Collection) and maintained in RPMI medium supplemented with 10% FBS. For efficacy studies, 5 × 106 Mia-Paca-2 cells were inoculated into the right posterior flank of 6- to 8-week-old BALB/c nude mice (Charles River Laboratories, Wilmington, MA, USA). Inoculate subcutaneously. Once the tumor volume is approximately 150 to 250 mm3, mice are randomized according to tumor volume and administered orally with the compound. Tumor volume is calculated using the formula (L×W2)/2, where L and W refer to the length and width dimensions, respectively. Tumor growth inhibition was calculated using the formula (1-(VT/VC))×100, where VT is the tumor volume of the treatment group on the last day of treatment and VC is the tumor volume of the control group on the last day of treatment. Two-way analysis of variance using Dunnett's multiple comparisons test is used to determine statistical differences between treatment groups (GraphPad Prism). Mice are housed at 10 to 12 animals per cage, fed well and exposed to a 12-hour light/dark cycle. Mice whose tumor volume exceeds the threshold (10% of body weight) are humanely euthanized by CO ₂ inhalation. Animals are maintained in barrier facilities fully accredited by the Association for Assessment and Accreditation of Laboratory Animal Care, International. All procedures are performed in accordance with the US Public Service Policy on Human Care and Use of Laboratory Animals and the Incyte Animal Care and Use Committee Guidelines.

Example H: Caco2 assay

Caco-2 cells were grown in DMEM growth medium supplemented with 10% (v/v) fetal bovine serum, 1% (v/v) non-essential amino acids, penicillin (100 U/mL), and streptomycin (100 μg/mL). It was grown at 37°C in a 5% CO ₂ atmosphere. Confluent cell monolayers were treated with 0.05% trypsin containing 1 μM EDTA and subcultured every 7 days or 4 days for Caco-2. Caco-2 cells were seeded in 96-well transwell plates. The seeding density for Caco-2 cells was 14,000 cells/well. DMEM growth medium was changed every other day after sowing. Cell monolayers were used for transport assays between days 22 and 25 for Caco-2 cells.

Cell culture medium was removed and replaced with HBSS. To measure TEER, HBSS was added to the donor compartment (apical side) and the recipient compartment (basal side). To ensure the integrity of the cell monolayer, TEER was measured using a REMS Autosampler. Caco-2 cell monolayers with TEER values ≥ 300 W·cm ² were used for transport experiments. To determine P _app in the absorption direction (AB), a solution of test compound (50 μM) in HBSS was added to the donor compartment (apical side), while a solution of HBSS with 4% BSA was added to the recipient compartment (basal side). Added. The apical volume was 0.075 mL and the basal volume was 0.25 mL. The incubation period was 120 minutes at 37°C in a 5% CO ₂ atmosphere. At the end of the incubation period, samples from the donor side and recipient side were removed and an equal volume of acetonitrile was added for protein precipitation. Supernatants were collected after centrifugation (3000 rpm, Allegra X-14R centrifuge from Beckman Coulter (Indianapolis, IN)) for LCMS analysis. Permeability values were determined according to the following equation:

P _app (cm/s) = (F * VD)/(SA * MD)

where the flux rate (F, mass/time) is calculated from the slope of the cumulative amount of the compound of interest on the recipient side, SA is the surface area of the cell membrane, VD is the donor volume, and MD is the initial volume of solution in the donor chamber. .

Caco-2 permeability assay data is provided in Table 3 below. The symbol “†” indicates Caco2 ≤ 0.5; And “††” indicates that Caco-2 is greater than 0.5 and less than or equal to 1; And “†††” indicates that Caco-2 is greater than 1. “NA” indicates that Caco-2 data is not available.

Caco-2 permeability assay data for certain compounds from WO 2021/142252 are provided in Table 4.

Example I: Human whole blood stability

Whole blood stability of the exemplified compounds was determined by LC-MS/MS. The 96-Well Flexi-Tier™ Block (Analytical Sales & Services, Inc, Flanders, NJ) contains 0.5 mL of blood (mixed gender, human whole blood or similar sourced from BIOIVT (Hicksville, NY)) per vial. It is used for incubation plates containing 1.0 mL glass vials. The blood is warmed to 37°C in a water bath for 30 minutes. A 96-deep well assay plate is prepared with the addition of 100 μL ultrapure water/well. Add 50 μL of chilled ultrapure water/well to a 96-deep well sample collection plate and cover with a sealing mat. Add 1 μL of 0.5 mM compound working solution (DMSO:water) to the blood in the incubation plate to reach a final concentration of 1 μM, mix thoroughly by pipetting, and then transfer 50 μL to the T=0 well of the sample collection plate. Blood is soaked in water for 2 minutes and then 400 μL of stop solution/well (acetonitrile containing internal standard) is added. The incubation plate is placed in an Incu-Shaker CO ₂ Mini incubator (Benchmark Scientific, Sayreville, NJ) at 37°C with shaking at 150 rpm. At 1, 2 and 4 hours, the blood samples are mixed thoroughly by pipetting and 50 μL is transferred to the corresponding well of the sample collection plate. After soaking the blood in water for 2 minutes, add 400 μL of stop solution/well. The collection plate was sealed and vortexed at 1700 rpm for 3 minutes (VX-2500 Multi-Tube Vortexer, VWR International, Lardner, PA), and samples were centrifuged in the collection plate at 3500 rpm for 10 minutes (Allegra X-14R Centrifuge). Beckman Coulter, Indianapolis, Indiana). Transfer 100 μL of supernatant/well from the sample collection plate to the corresponding well of the assay plate. The final plate is vortexed at 1700 rpm for 1 minute and samples are analyzed by LC-MS/MS. The peak area ratios of the 1, 2 and 4 hour samples to T=0 are used to determine the percentage remaining. Calculate the compound half-life in blood (t _1/2 = 0.693/slope) by determining the slope using the natural logarithm of percentage remaining versus time.

Human whole blood stability data is provided in Table 5 below. The symbol “†” indicates WBS ≤ 70%; “††” indicates WBS is greater than 70% but less than 90%; And “†††” indicates WBS > 90%. “NA” indicates that WBS data is not available.

Example J: In vitro endogenous clearance protocol

For in vitro metabolic stability experiments, test compounds are incubated with human liver microsomes at 37°C. The incubation mixture contains test compound (1 μM), NADPH (2 mM) and human liver microsomes (0.5 mg protein/mL) in 100 mM phosphate buffer (pH 7.4). This mixture is preincubated at 37°C for 2 minutes before adding NADPH. The reaction starts when NADPH is added, and the reaction is stopped with ice-cold methanol at 0, 10, 20, and 30 minutes. The finished incubation mixture is analyzed using an LC-MS/MS system. The analysis system consisted of a Shimadzu LC-30AD binary pump system and SIL-30AC autosampler (Shimadzu Scientific Instruments, Columbia, MD) coupled to a Sciex Triple Quad 6500+ mass spectrometer from Applied Biosystems (Foster City, CA). . Chromatographic separation of test compounds and internal standards is achieved using a Hypersil Gold C18 column (50×2.1 mm, 5 μM, 175 Å) from ThermoFisher Scientific (Waltham, MA). Mobile phase A consists of 0.1% formic acid in water, and mobile phase B consists of 0.1% formic acid in acetonitrile. The total LC-MS/MS run time can be 2.75 minutes with a flow rate of 0.75 mL/min. Peak area integration and peak area ratio calculations are performed using Analyst software (version 1.6.3) from Applied Biosystems.

The in vitro endogenous clearance rate, CL _{int, in vitro} , is calculated from t _1/2 of the disappearance of the test compound as CL _{int, in vitro} = (0.693/ t _1/2 ) × (1/ C _protein ), where C _protein is is the protein concentration during, and t _1/2 is determined by the slope (k) of a log-linear regression of the concentration versus time profile; Therefore, t _1/2 =ln2/k. CL _{int, in vitro} values are scaled to in vivo values for humans using physiologically based scaling factors: liver microsomal protein concentration (45 mg protein/g liver) and liver weight (21 g/kg body weight). The equation CL _int =CL _{int, in vitro} × (mg protein/g liver weight) × (g liver weight/kg body weight) is used. Then, in vivo Hepatic clearance (CL _H ) is calculated as CL _int and hepatic blood flow Q (20 mL min in humans) in a well-stirred ^liver model, ignoring all coupling from CL _H = (QХCL _int )/(Q+CL _int It is calculated using ¹ kg ^-1 ). The liver extraction ratio was calculated by dividing CL _H by Q.

Example K: In Vivo Pharmacokinetic Protocol

For in vivo pharmacokinetic experiments, test compounds are administered intravenously or via oral gavage to male Sprague Dawley rats or male and female Cynomolgus monkeys. For intravenous (IV) administration, test compounds are administered in a formulation of 10% dimethylacetamide (DMAC) in acidified saline via IV bolus in rats and via 5- or 10-minute IV infusion in monkeys. Administered at 0.5 to 1 mg/kg. For oral (PO) administration, test compounds are administered at 1.0 to 3.0 mg/kg using 5% DMAC in 0.5% methylcellulose in citrate buffer (pH 2.5). Blood samples are collected pre-dose and at various time points up to 24 hours post-dose. All blood samples are collected using EDTA as an anticoagulant and centrifuged to obtain plasma samples. Plasma concentrations of test compounds are determined by LC-MS method. The measured plasma concentrations are used to calculate PK parameters by standard non-compartmental methods using the ^Phoenix® WinNonlin software program (version 8.0, Pharsight Corporation).

In rats and monkeys, cassette administration of test compounds is performed to obtain preliminary PK parameters.

In vivo pharmacokinetic experiments with male beagle dogs can be performed under the conditions described above.

Example L: Time-dependent inhibition (TDI) of the CYP protocol

This assay is designed to characterize the increase in CYP inhibition as the test compound is metabolized over time. Potential mechanisms for this include inactivation of P450 enzymes by tight binding, formation of semireversible inhibitory metabolite complexes, or covalent adduct formation of metabolites. Although this experiment uses a 10-fold dilution to reduce the metabolite concentration and therefore the reversible inhibitory effect, it is possible (but not common) that metabolites that are very strong CYP inhibitors may produce positive results.

The results are derived from a cocktail of CYP-specific probe substrates at four times the Km concentration for CYP2C9, 2C19, 2D6 and 3A4 (midazolam) using human liver microsomes (HLM). HLMs were preincubated with test compounds at a concentration of 10 μM for 30 min in the presence (+N) or absence (-N) of the NADPH regeneration system, diluted 10-fold, and the substrate cocktail with the addition of a fresh aliquot of the NADPH regeneration system. can be incubated for 8 minutes in the presence of Enzyme activity can be measured quantitatively using LC-MS/MS using a calibration curve of metabolite standards. Additionally, incubation with the known time-dependent inhibitors used as positive controls, thienyl acid (CYP2C9), ticlopidine (CYP2C19), paroxetine (CYP2D6), and troleandomycin (CYP3A4), was performed for 30 min with or without the NADPH regeneration system. Preliminary incubation is performed.

The analysis system consists of a Shimadzu LC-30AD binary pump system and a SIL-30AC autosampler (Shimadzu Scientific Instruments, Columbia, MD) coupled to a Sciex Triple Quad 6500+ mass spectrometer from Applied Biosystems (Foster City, CA). Chromatographic separation of test compounds and internal standards can be achieved using an ACQUITY UPLC BEH 130A, 2.1×50 mm, 1.7 μm HPLC column (Waters Corp, Milford, MA). Mobile phase A consists of 0.1% formic acid in water, and mobile phase B consists of 0.1% formic acid in acetonitrile. Total LC-MS/MS run time will be 2.50 minutes with a flow rate of 0.9 mL/min. Peak area integration and peak area ratio calculations are performed using Analyst software (version 1.6.3) from Applied Biosystems.

The percentage of control CYP2C9, CYP2C19, CYP2D6 and CYP3A4 activity remaining after preincubation of compounds containing NADPH is corrected for the corresponding control vehicle activity and then calculated relative to 0 minutes as 100%. For each isoenzyme, the slope is calculated using a linear regression plot of the natural logarithm of % activity remaining versus time. -The slope is equivalent to the enzyme loss rate or K _obs .

Various modifications of the present invention in addition to those described herein will become apparent to those skilled in the art from the above description. Such modifications are also intended to be included within the scope of the appended claims. Each reference, including all patents, patent applications and publications, cited in this application is hereby incorporated by reference in its entirety.

Claims (33)

A compound of formula (I) or a pharmaceutically acceptable salt thereof:

During the ceremony:
Y is N or CH;
R ¹ is selected from Cl, CH ₃ , CH ₂ F, CHF ₂ and CF ₃ ;
Cy ¹ is selected from:


R ² is selected from F and Cl;
R ³ is selected from:

and
Cy ² is selected from:

However, the compounds of formula I are other than:
The compound according to claim 1, wherein the compound of formula (I) is a compound of formula (II) or a pharmaceutically acceptable salt thereof:

During the ceremony:
R ¹ is selected from Cl and CH ₃
Cy ¹ is selected from:

R ³ is selected from:

and,
Cy ² is selected from:
According to paragraph 1,
Y is N or CH;
R ¹ is selected from Cl, CH ₃ , CH ₂ F, CHF ₂ and CF ₃ ;
Cy ¹ is selected from:

R ² is selected from F and Cl;
R ³ is selected from:

and
Cy ² is selected from:

provided that the compound of formula (I) is other than:
According to claim 1 or 3,
Y is N or CH;
R ¹ is selected from Cl, CH ₃ , CH ₂ F, CHF ₂ and CF ₃ ;
Cy ¹ is selected from:

R ² is selected from F and Cl;
R ³ is selected from:

and,
Cy ² is a compound or a pharmaceutically acceptable salt thereof selected from:
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein Y is CH. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein Y is N. The compound or pharmaceutical composition according to any one of claims 1 to 6, wherein Cy ¹ is selected from Cy ¹ -a, Cy ¹ -b, Cy ¹ -c, Cy ¹ -d and Cy ¹ -e. Acceptable salts. The method according to any one of claims 1 to 6, wherein Cy ¹ is selected from Cy ¹ -f, Cy ¹ -g, Cy ¹ -h, Cy ¹ -i, Cy ¹ -j and Cy ¹ -k. A compound or a pharmaceutically acceptable salt thereof. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein Cy ¹ is selected from Cy ¹ -a and Cy ¹ -b. The method of claim 1 or 2, wherein Cy ¹ is Cy ¹ -l, Cy ¹ -m, Cy ¹ -n, Cy ¹ -o, Cy ¹ -p, Cy ¹ -q, Cy ¹ -r, Cy ¹ A compound or a pharmaceutically acceptable salt thereof selected from -s and Cy ¹ -t. The method of claim 1 or 2, wherein Cy ¹ is Cy ¹ -c, Cy ¹ -m, Cy ¹ -n, Cy ^{1 -o, Cy 1 -p} , Cy ¹ -q, Cy ¹ -r, Cy ¹ A compound or a pharmaceutically acceptable salt thereof selected from -s and Cy ¹ -t. 12. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, wherein R ¹ is selected from CH ₃ , CH ₂ F, CHF ₂ and CF ₃ . 13. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 12, wherein R ¹ is selected from CH ₃ and CF ₃ . The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 and 3 to 13, wherein R ² is F. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 and 3 to 13, wherein R ² is Cl. 16. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 15, wherein R ³ is selected from R ³ -a and R ³ -b. 16. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 and 3 to 15, wherein R ³ is selected from R ³ -b and R ³ -c. 18. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 and 3 to 17, wherein Cy ² is selected from Cy ² -a, Cy ² -b and Cy ² -d. 19. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 18, wherein Cy ² is selected from Cy ² -a and Cy ² -b. 19. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 18, wherein Cy ² is selected from Cy ² -b and Cy ² -d. 4. Compound according to claim 1 or 3, wherein the compound of formula I is selected from:
2-((2 S ,4 S )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinoline- 8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4- fluorobut-2-enoyl)piperidin-2-yl)acetonitrile;
2-((2 S ,4 S )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(2-methoxy-3 -methylphenyl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4-fluo Robut-2-enoyl)piperidin-2-yl)acetonitrile;
2-((2 S ,4 S )-4-(7-(3-chloro-2-methoxyphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)- 6-fluoro-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4- fluorobut-2-enoyl)piperidin-2-yl)acetonitrile;
2-(( 2S , 4S )-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-(( S )-1-(( S )-1-methyl Pyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1- (( E )-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile;
1-(4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-(( S )-1-(( S )-1-methylpyrrolidin-2-yl) Ethoxy)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)piperidin-1-yl)prop- 2-en-1-one;
2-((2 S ,4 S )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluo Ro-8-methyl- 1H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4-methoxybut-2-enoyl )piperidin-2-yl)acetonitrile;
2-(( 2S , 4S )-4-(6-fluoro-8-methyl-7-(1-methyl- 1H -indazol-6-yl)-4-(( S )-1- (( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-( ( E )-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile;
2-(( 2S , 4S )-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-(( S )-1-(( S )- 1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4 -fluorobut-2-enoyl)piperidin-2-yl)acetonitrile;
2-(( 2S , 4S )-4-(6-fluoro-8-methyl-7-(1-methyl- 1H -indazol-3-yl)-4-(( S )-1- (( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-( ( E )-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile;
2-(( 2S , 4S )-4-(6-fluoro-7-(4-fluorophenyl)-8-methyl-4-(( S )-1-(( S )-1-methyl Pyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4-methoxy but-2-enoyl)piperidin-2-yl)acetonitrile;
8-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-(( S )-1-(( S )-1-methylpyrrolidin-2 -yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)-1-naphtonitrile;
2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro -8-methyl-1H-imidazo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)aceto nitrile;
2-((2 S ,4 S )-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-(( S )-1-(( S )-1 -methylpyrrolidin-2-yl)ethoxy)-1 H -imidazo[4,5- c ]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidine-2- 1) Acetonitrile;
8-(1-((2 R ,4 S )-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-(( S )-1- (( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)-1-naph Tonitrile;
2-((2 S ,4 S )-4-(7-(5,6-dimethyl-1 H -indazol-4-yl)-4-(3-(ethyl(methyl)amino)azetidine- 1-yl)-6-fluoro-8-methyl-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(2-fluoroacrylo 1) piperidin-2-yl) acetonitrile;
8-(6-fluoro-1-(1-(( E )-4-fluorobut-2-enoyl)piperidin-4-yl)-8-methyl-4-(( S )-1 -(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)-1- naphtonitrile;
8-(1-((2 S ,4 S )-2-(cyanomethyl)-1-(2-fluoroacryloyl)piperidin-4-yl)-4-(3-(dimethyl amino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)- 1-naphtonitrile;
2-((2 S ,4 S )-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoro Quinolin-8-yl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4-methoxybut-2-ene oil) piperidin-2-yl) acetonitrile;
2-((2 S ,4 S )-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoro Quinolin-8-yl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4-fluorobut-2-ene oil) piperidin-2-yl) acetonitrile; and
2-((2 S ,4 S )-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinoline- 8-yl)-8-(trifluoromethyl)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-1-yl)-1-(( E )-4- methoxybut-2-enoyl)piperidin-2-yl)acetonitrile;
Or a pharmaceutically acceptable salt thereof. 2. The compound according to claim 1, wherein the compound of formula I is selected from:
2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-((S)- 1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4- methoxybut-2-enoyl)piperidin-2-yl)acetonitrile;
2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-((S)- 1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacrylo 1) piperidin-2-yl) acetonitrile;
2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1 -methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl )piperidin-2-yl)acetonitrile;
2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1 -methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-meth Toxybut-2-enoyl)piperidin-2-yl)acetonitrile;
2-((2S,4S)-4-(6-fluoro-7-(7-fluoro-2-methylquinolin-8-yl)-8-methyl-4-((S)-1-(( S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluo Roacryloyl)piperidin-2-yl)acetonitrile;
2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methylisoquinolin-4-yl)-4-((S)-1-((S)-1 -methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-meth Toxybut-2-enoyl)piperidin-2-yl)acetonitrile;
2-((2S,4S)-4-(6-fluoro-7-(7-fluoroquinolin-8-yl)-8-methyl-4-((S)-1-((S)-1 -methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl )piperidin-2-yl)acetonitrile;
2-(4-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1 -((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl -1H-indazol-3-yl)acetonitrile;
2-((2S,4S)-4-(4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5 -methyl-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoro Acryloyl)piperidin-2-yl)acetonitrile;
2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidine -1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacrylo 1) piperidin-2-yl) acetonitrile;
2-(4-(1-((2 R ,4 S )-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-(( S ) -1-(( S )-1-methylpyrrolidin-2-yl)ethoxy)-1 H -[1,2,3]triazolo[4,5- c ]quinolin-7-yl)- 1-methyl-1 H -indol-3-yl)acetonitrile;
2-(4-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-(( S )-1-(( S )-1-methylpyrroli din-2-yl) ethoxy) -1 H -[1,2,3] triazolo [4,5- c ] quinolin-7-yl) -1-methyl-1 H -indol-3-yl) acetonitrile;
2-((2S,4S)-4-(8-chloro-7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3 -methylazetidin-1-yl)-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacrylo 1) piperidin-2-yl) acetonitrile;
2-((2S,4S)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-8-methyl-4-((S)-1-((S)- 1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacrylo 1) piperidin-2-yl) acetonitrile; and
2-((2S,4S)-4-(8-chloro-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6- Fluoro-5-methyl-1H-indazol-4-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoro Acryloyl)piperidin-2-yl)acetonitrile;
Or a pharmaceutically acceptable salt thereof. A pharmaceutical composition comprising the compound of any one of claims 1 to 22 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier or excipient. A method of inhibiting KRAS activity, comprising contacting KRAS with the compound of any one of claims 1 to 22 or the composition of claim 23. 25. The method of claim 24, wherein said linking step comprises administering said compound to the patient. A method of treating a disease or disorder associated with inhibition of KRAS interaction, comprising administering to a patient in need of treatment the disease or disorder a therapeutically effective amount of the compound of any one of claims 1 to 22 or the composition of claim 23. A method comprising administering. 1. A method of treating a disease or disorder associated with inhibiting a KRAS protein carrying a G12C mutation, comprising: providing a therapeutically effective amount of the compound of any one of claims 1 to 22 to a patient in need of treatment for the disease or disorder; Or a method comprising administering the composition of claim 23. A method of treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of the compound of any one of claims 1 to 22 or the composition of claim 23. 29. The method of claim 28, wherein the cancer is selected from carcinoma, hematological cancer, sarcoma, and glioblastoma. 30. The method of claim 29, wherein the hematologic malignancy is selected from myeloproliferative neoplasms, myelodysplastic syndromes, chronic and childhood myelomonocytic leukemias, acute myeloid leukemias, acute lymphoblastic leukemias, and multiple myeloma. 30. The method of claim 29, wherein the carcinoma is selected from pancreas, colorectal, lung, bladder, stomach, esophagus, breast, head and neck, cervix, skin and thyroid. 28. The method of claim 27, wherein the disease or disorder is an immune or inflammatory disorder. 33. The method of claim 32, wherein the immune or inflammatory disorder is Ras-associated lymphoproliferative disorder and pediatric myelomonocytic leukemia caused by somatic mutations in KRAS.